LOCUS       sequence-418201-        4923 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Neuronal expression of EGFP tagged KASH protein domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4923)
  TITLE     Zweifel lab KASH plasmids
REFERENCE   2  (bases 1 to 4923)
  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"
COMMENT     Sequence Label: pAAV-hSyn-EGFP-KASH
FEATURES             Location/Qualifiers
     source          1..4923
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        152..599
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      631..640
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(637..639,640..642,643..1353)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(637..639,640..642,643..1353)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(682..703)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(943..962)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1290..1311
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1670..2146
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1869..1888)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2186..2326
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2186..2315
                     /label=AAV2 ITR
     rep_origin      2401..2856
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2488..2507)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2698..2719
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2873..2892)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2992..3014
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3052..3070)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3138..3242
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3243..3311,3312..4103)
                     /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(3461..3480)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4274..4862
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4763..4782
                     /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 gcctgcgtat gagtgcaagt gggttttagg accaggatga
      181 ggcggggtgg gggtgcctac ctgacgaccg accccgaccc actggacaag cacccaaccc
      241 ccattcccca aattgcgcat cccctatcag agagggggag gggaaacagg atgcggcgag
      301 gcgcgtgcgc actgccagct tcagcaccgc ggacagtgcc ttcgcccccg cctggcggcg
      361 cgcgccaccg ccgcctcagc actgaaggcg cgctgacgtc actcgccggt cccccgcaaa
      421 ctccccttcc cggccacctt ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc
      481 acgcgaggcg cgagataggg gggcacgggc gcgaccatct gcgctgcggc gccggcgact
      541 cagcgctgcc tcagtctgcg gtgggcagcg gaggagtcgt gtcgtgcctg agagcgcagt
      601 cgagaaggta ccggatcctc tagagtcgac gccaccatgg tgagcaaggg cgaggagctg
      661 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
      721 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
      781 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
      841 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
      901 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
      961 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     1021 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     1081 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     1141 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     1201 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     1261 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     1321 gggatcactc tcggcatgga cgagctgtac aagtccggac tcagatctcg agaggaggag
     1381 gaggagacag acagcaggat gccccacctc gacagccccg gcagctccca gccgagacgc
     1441 tccttcctct caagggtgat cagggcagcg ctaccgttgc agctgcttct gctgctgctg
     1501 ctgctcctgg cctgcctgct acctgcctct gaagatgact acagctgcac ccaggccaac
     1561 aactttgccc gatccttcta ccccatgctg cggtacacca acgggccacc tcccacctag
     1621 gaattcgata tcaagcttat cgataccgag cgctgctcga gagatctacg ggtggcatcc
     1681 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag
     1741 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat
     1801 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc
     1861 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat
     1921 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca
     1981 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat
     2041 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa
     2101 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt
     2161 aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     2221 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     2281 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     2341 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     2401 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2461 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2521 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2581 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     2641 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     2701 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     2761 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     2821 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     2881 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     2941 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     3001 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     3061 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     3121 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3181 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3241 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3301 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3361 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     3421 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     3481 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     3541 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     3601 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     3661 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     3721 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     3781 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     3841 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     3901 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     3961 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     4021 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4081 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     4141 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4201 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4261 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4321 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4381 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     4441 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4501 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4561 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4621 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     4681 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4741 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4801 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4861 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4921 tgt
//
