LOCUS       sequence-352277-        5367 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV transfer plasmid for cell-surface-localized FRISZ (via PDGFR TM)
            under hSyn promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5367)
  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 5367)
  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-TM
FEATURES             Location/Qualifiers
     source          1..5367
                     /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"
     sig_peptide     647..706
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             1550..1579
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1583..1729
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    1757..2345
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1810..1830)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2228..2239)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2387..2404)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2393..2600
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2629..2769
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2629..2758
                     /label=AAV2 ITR
     rep_origin      2844..3299
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2931..2950)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3141..3162
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3316..3335)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3435..3457
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3495..3513)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3581..3685
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3686..3754,3755..4546)
                     /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(3904..3923)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4717..5305
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5206..5225
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(5367,1..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 agacagacac
      661 actcctgcta tgggtactgc tgctctgggt tccaggttcc actggtgaca gatctatggt
      721 agatgcaaag ggaaaacacc cagtacacgg tgctgaatta acagatgtgt ccaagaaacc
      781 cgcaaagaac ctcaagatgc caggcgtcta ttttattgac aggattctgg aaaggatcaa
      841 ggaggctggc aatgagacct acgtcgagca gtacgaggtg gctgtggcca gatactgcga
      901 ccaccctagc aaactggggc acaaacttaa aggaggaaca ggaggaagca gcgaggagct
      961 gatcaaggag aacatgcgca ccaagctgta cctgacgggc accgtgaaca accactactt
     1021 cgaatgcacc gccgaagggg agggccagcc ctatgagggc acccagacca acaggattaa
     1081 ggtggtgcgc ggaggccccc tgccgttcgc attcgacatc ctggccccct gctttatgta
     1141 cgggagcaag accttcatta accatccacc tgacattccc gattatttta agcagtcctt
     1201 ccctgagggc ttcacatggg agagaaccac cgttttcgaa gacgggggca cactcactgc
     1261 tatacaggac accagcctcc aggacggctg cctgatctac aacgtccaag tcagagggga
     1321 gaacttccca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg aggccagtac
     1381 cgagaccctg taccccgctg acggcagcct ggaaggcaga ctgtactggg ccctgaagct
     1441 cgtgggcggg ggccacctgc actgcagact ggaaaccaca tacagatcca agaaattgag
     1501 tgctaaaggt aagcatccag tacatggagc agagctgact gacgtcgacg aacaaaaact
     1561 catctcagaa gaggatctga atgctgtggg ccaggacacg caggaggtca tcgtggtgcc
     1621 acactccttg ccctttaagg tggtggtgat ctcagccatc ctggccctgg tggtgctcac
     1681 catcatctcc cttatcatcc tcatcatgct ttggcagaag aagccacgtt aagaattcga
     1741 tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     1801 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     1861 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     1921 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     1981 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     2041 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     2101 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     2161 ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt ccttctgcta
     2221 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     2281 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     2341 cccgcatcga taccgagcgc tgctcgagct agagctcgct gatcagcctc gactgtgcct
     2401 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     2461 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2521 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     2581 aatagcaggc atgctgggga ggtaaccacg tgcggaccga gcggccgcag gaacccctag
     2641 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     2701 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     2761 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     2821 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2881 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     2941 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3001 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     3061 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3121 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     3181 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     3241 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     3301 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     3361 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3421 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3481 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     3541 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3601 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3661 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     3721 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     3781 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     3841 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     3901 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     3961 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4021 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4081 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4141 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4201 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4261 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4321 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4381 atctggagcc ggtgagcgtg gaagccgcgg tatcattgca gcactggggc cagatggtaa
     4441 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4501 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4561 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4621 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4681 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4741 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4801 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4861 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4921 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4981 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5041 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5101 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5161 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5221 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5281 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5341 cttttgctgg ccttttgctc acatgtc
//
