LOCUS       sequence-352276-        5339 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV transfer plasmid for cell-surface-localized FRISZ (via CD59 GPI)
            under hSyn promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5339)
  AUTHORS   Wu T, Kumar M, Zhang J, Zhao S, Drobizhev M, McCollum M, Anderson 
            CT, Wang Y, Pokorny A, Tian X, Zhang Y, Tzounopoulos T, Ai HW
  TITLE     A genetically encoded far-red fluorescent indicator for imaging 
            synaptically released Zn(2).
  JOURNAL   Sci Adv. 2023 Mar;9(9):eadd2058. doi: 10.1126/sciadv.add2058. Epub 
            2023 Mar 1.
  PUBMED    36857451
REFERENCE   2  (bases 1 to 5339)
  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: "Sci Adv."; 
            date: "2023-03"; volume: "9(9):eadd2058. doi"; pages: " 
            10.1126/sciadv.add2058. Epub 2023 Mar 1"
COMMENT     Sequence Label: pAAV-hSyn-FRISZ-GPI
FEATURES             Location/Qualifiers
     source          1..5339
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        180..627
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      641..650
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1729..2317
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1782..1802)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2200..2211)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2359..2376)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2365..2572
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2601..2741
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2601..2730
                     /label=AAV2 ITR
     rep_origin      2816..3271
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2903..2922)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3113..3134
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3288..3307)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3407..3429
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3467..3485)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3553..3657
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3658..3726,3727..4518)
                     /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(3876..3895)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4689..5277
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5178..5197
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(5339,1..129)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcgtcgg gcgacctttg
       61 gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta
      121 ggggttcctg cggccgcacg cgttgtacag tgtctagact gcagagggcc ctgcgtatga
      181 gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct gacgaccgac
      241 cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag
      301 agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg
      361 acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg
      421 ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc
      481 gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc
      541 gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga
      601 ggagtcgtgt cgtgcctgag agcgcagtcg agaaggatcc gccaccatgg gaatccaagg
      661 agggtctgtc ctgttcgggc tgctgctcgt cctggctgtc ttctgccatt caggtcatag
      721 cagatctatg gtcgacgcaa agggaaaaca cccagtacac ggtgctgaat taacagatgt
      781 gtccaagaaa cccgcaaaga acctcaagat gccaggcgtc tattttattg acaggattct
      841 ggaaaggatc aaggaggctg gcaatgagac ctacgtcgag cagtacgagg tggctgtggc
      901 cagatactgc gaccacccta gcaaactggg gcacaaactt aaaggaggaa caggaggaag
      961 cagcgaggag ctgatcaagg agaacatgcg caccaagctg tacctgacgg gcaccgtgaa
     1021 caaccactac ttcgaatgca ccgccgaagg ggagggccag ccctatgagg gcacccagac
     1081 caacaggatt aaggtggtgc gcggaggccc cctgccgttc gcattcgaca tcctggcccc
     1141 ctgctttatg tacgggagca agaccttcat taaccatcca cctgacattc ccgattattt
     1201 taagcagtcc ttccctgagg gcttcacatg ggagagaacc accgttttcg aagacggggg
     1261 cacactcact gctatacagg acaccagcct ccaggacggc tgcctgatct acaacgtcca
     1321 agtcagaggg gagaacttcc catccaacgg ccctgtgatg cagaagaaaa cactcggctg
     1381 ggaggccagt accgagaccc tgtaccccgc tgacggcagc ctggaaggca gactgtactg
     1441 ggccctgaag ctcgtgggcg ggggccacct gcactgcaga ctggaaacca catacagatc
     1501 caagaaattg agtgctaaag gtaagcatcc agtacatgga gcagagctga ctgacggctc
     1561 cacctccggc tccggcaagc ccggctccgg cgagggctcc accaagggcg aattccttga
     1621 aaatggtggg acatccttat cagagaaaac agttcttctg ctggtgactc catttctggc
     1681 agcagcctgg agccttcatc cctaagaatt catatcaagc ttatcgataa tcaacctctg
     1741 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     1801 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     1861 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     1921 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     1981 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     2041 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     2101 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc
     2161 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     2221 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     2281 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag
     2341 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     2401 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2461 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     2521 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gaggtaacca
     2581 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     2641 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     2701 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     2761 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     2821 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2881 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2941 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3001 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     3061 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3121 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
     3181 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3241 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     3301 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     3361 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     3421 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     3481 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     3541 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3601 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     3661 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     3721 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     3781 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     3841 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     3901 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     3961 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4021 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4081 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4141 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     4201 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     4261 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     4321 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tggaagccgc
     4381 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     4441 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     4501 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     4561 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4621 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4681 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4741 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4801 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     4861 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4921 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4981 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5041 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5101 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5161 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5221 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5281 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtc
//
