LOCUS       sequence-495225-        5570 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for packing AAV for hpGRISZ (synaptic zinc sensor).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5570)
  AUTHORS   Zhang H, Kumar M, Correia R, Yang S, Huang W, Soares-Cunha C, Cody 
            PA, Zhang Y, Tian X, Zhao S, Drobizhev M, Rodrigues AJ, Tzounopoulos
            T, Ai H
  TITLE     hpGRISZ: a high-performance fluorescent biosensor for in vivo 
            imaging of synaptic zinc dynamics
  JOURNAL   bioRxiv 2025.10.06.680726
REFERENCE   2  (bases 1 to 5570)
  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: "bioRxiv 
            2025.10.06.680726"
COMMENT     Sequence Label: pAAV-hSyn-hpGRISZ-TM
FEATURES             Location/Qualifiers
     source          1..5570
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     190..209
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(428..532)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(558..1013)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(695..716)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     907..926
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1085..1214
                     /label=AAV2 ITR
     promoter        1290..1737
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      1766..1775
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1772..1831
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             2897..2926
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2930..3076
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    3092..3680
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3145..3165)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3563..3574)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3683..3699)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3684..3700)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3729..3850
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3740..3759)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3794..3813
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4012..4141
                     /label=AAV2 ITR
     rep_origin      complement(4378..4966)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4458..4477)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5137..5570,1..358,359..427))
                     /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"
ORIGIN
        1 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
       61 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
      121 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
      181 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
      241 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
      301 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
      361 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
      421 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
      481 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
      541 cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
      601 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
      661 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
      721 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
      781 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
      841 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
      901 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
      961 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc
     1021 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     1081 ccagctgcgc gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac
     1141 ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat
     1201 cactaggggt tccttgtagt taatgattaa cccgccatgc tacttatcta cgtagccatg
     1261 ctctaggaag atccagggcc ctgcgtatga gtgcaagtgg gttttaggac caggatgagg
     1321 cggggtgggg gtgcctacct gacgaccggc cccgacccac tggacaagca cccaaccccc
     1381 attccccaaa ttgcgcatcc cctatcagag agggggaggg gaaacaggat gcggcgaggc
     1441 gcgtgcgcac tgccagcttc agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg
     1501 cgccaccgcc gcctcagcac tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact
     1561 ccccttcccg gccaccttgg tcgcgtccgc gccgccgccg gcccagccgg accgcaccac
     1621 gcgaggcgcg agataggggg gcacgggcgc gaccatctgc gctgcggcgc cggcgactca
     1681 gcgctgcctc agtctgcggt gggcagcgga ggagtcgtgt cgtgcctgag agcgcagtcc
     1741 aattcaagct gctagcaagg atcccgccac catggagaca gacacactcc tgctatgggt
     1801 actgctgctc tgggttccag gttccactgg tgacagatct atgaacaata acgatctctt
     1861 tcaggcatca cgtcggcgtt ttctggcaca actcggcagc ttaaccgacg ccgggacgct
     1921 ggggccgtca ttgttaacgc cgcgacgtgc gactgcggcg caagcggcga ctgacgcttc
     1981 tagaggtatg gctggcgtga ctggtggaca gcaaatgggt cgggatctgt acgacgatga
     2041 tgataatgat ctcgccactc tcgagttggt agacgctaag agaaaacacc cagtacatgg
     2101 tgttgaatta acagattatg tttatatcaa ggccgacaag cggaggaatg gcatcaaggc
     2161 gaacttcctg atccgccaca acgtggagga cggcagcgtg cagctcgccg accactacca
     2221 gcggaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgatgta
     2281 ccagaccgtg ctgagcaaag accccaacga gaagcgagat cacatggtcc tgctggagta
     2341 cgtgaccgcc gccgggatta ctctcggcgg tagtgggggg caaagggtgg acggtggatc
     2401 cggtggcacc ggggtgagca ggggcgagga gctgttcacc ggggtggtgc ccatcctggt
     2461 cgtactggac ggcgtcgtta acggccacaa gttccgcgtg aggggcgaag gtgagggcga
     2521 tgccaccaac ggcaagctga ccttgaagtt catctgcact accggcatgc tgccagtgcc
     2581 ctggcccacc ctcgtgacca cctttaccta cggcgtgcag tgcttcgccc ggtaccccga
     2641 ccacatgaag cagcacgact tcttcaagtc cgcgatgccc gaaggctaca tccaggagcg
     2701 caacatcttc ttcaaggacg acggcaccta caagacccgc gccgaggtga agttcgaggg
     2761 cgacaccctg gtgaaccgca tcgagctgaa gggcgtcgac ttcaaggagg atggcaacat
     2821 cctggggcac aagctggagt tcgagtttgc taaaggtgag catccagtac atggagcaaa
     2881 gctgactgac gtcgacgaac aaaaactcat ctcagaagag gatctgaatg ctgtgggcca
     2941 ggacacgcag gaggtcatcg tggtgccaca ctccttgccc tttaaggtgg tggtgatctc
     3001 agccatcctg gccctggtgg tgctcaccat catctccctt atcatcctca tcatgctttg
     3061 gcagaagaag ccacgttaga agcttatcga taatcaacct ctggattaca aaatttgtga
     3121 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     3181 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     3241 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     3301 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     3361 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     3421 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     3481 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     3541 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     3601 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     3661 cctttgggcc gcctccccgc atcgataccg tcgacctcga cccgggcggc cgcttcgagc
     3721 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     3781 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     3841 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg
     3901 ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatcttcc
     3961 tagagcatgg ctacgtagat aagtagcatg gcgggttaat cattaactac aaggaacccc
     4021 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     4081 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     4141 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4201 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4261 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4321 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4381 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4441 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4501 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4561 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4621 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4681 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4741 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4801 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4861 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4921 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4981 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5041 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5101 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5161 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5221 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5281 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5341 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5401 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5461 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5521 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
//
