LOCUS       sequence-461237-        6920 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV encoding PiGM-Iq with minimal RGS10 domain, CRY2 (535 amino acid
            version), CIB1 (81 amino acid version)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6920)
  TITLE     PiGM-Iq system plasmids
REFERENCE   2  (bases 1 to 6920)
  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: AAV-hSyn-PiGM-Iq(535/81, eGFP)
FEATURES             Location/Qualifiers
     source          1..6920
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     294..313
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(532..636)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(662..1117)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(799..820)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1011..1030
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1189..1318
                     /label=AAV2 ITR
     promoter        1400..1847
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             4019..4072
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(4364..4366,4367..4369,4370..5080)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4364..4366,4367..4369,4370..5080)
                     /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(4409..4430)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4670..4689)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5017..5038
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5235..5255)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     rep_origin      complement(5899..6487)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5979..5998)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
       61 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
      121 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
      181 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
      241 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
      301 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
      361 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
      421 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
      481 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
      541 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
      601 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
      661 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
      721 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
      781 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
      841 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
      901 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
      961 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     1021 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     1081 acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc
     1141 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct gcgcgctcgc
     1201 tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc
     1261 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttccttg
     1321 tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag gaagatctct
     1381 gcagagggcc ctgcgtatga gtgcaagtgg gttttaggac caggatgagg cggggtgggg
     1441 gtgcctacct gacgaccgac cccgacccac tggacaagca cccaaccccc attccccaaa
     1501 ttgcgcatcc cctatcagag agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac
     1561 tgccagcttc agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc
     1621 gcctcagcac tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg
     1681 gccaccttgg tcgcgtccgc gccgccgccg gcccagccgg accgcaccac gcgaggcgcg
     1741 agataggggg gcacgggcgc gaccatctgc gctgcggcgc cggcgactca gcgctgcctc
     1801 agtctgcggt gggcagcgga ggagtcgtgt cgtgcctgag agcgcagtcg aattcaagct
     1861 tggatccatg actcctggga agcccaaaac tggcaagaaa agcaaacagc aaacttttat
     1921 caagccttct cctgaggaag cgctgctctg ggcagaagca tttgatgaac tgctggccag
     1981 caaatatggg cttgctgcat tcagagcgtt tttaaagtcc gagttctgtg aagaaaatat
     2041 tgaattctgg ttggcttgcg aagacttcaa gaaaaccaaa tcaccacaga aactgtcctc
     2101 aaaagcaagg aaaatataca ccgacttcat cgagaaggaa gctcccaaag agataaacat
     2161 agactttcaa acaaaaactc tgattgccca aaatatccaa gaggctacaa gtggctgctt
     2221 caccacagcg cagaagaggg tgtacagcct gatggagaac aactcttatc ctcgtttctt
     2281 ggaatcagaa ttttatcagg acttgtgtaa aaagccacag atcaccacgg agccccatgc
     2341 tacaaccggt ggcggcagtg gtggcggcag catgaagatg gacaagaaga caattgtctg
     2401 gtttcgcagg gacctgagaa tcgaagataa ccccgcattg gcagcagcag cccatgaggg
     2461 ttctgtgttc cctgtcttca tttggtgtcc agaggaggag ggacagttct accctggccg
     2521 ggccagcaga tggtggatga agcagtcctt ggctcacctg tcacagagtc tcaaggcact
     2581 gggctccgat ctgacgttga ttaagaccca caacacgatt agcgcaattc tggactgcat
     2641 ccgggtgacc ggcgcaacaa aagtggtttt caaccacctc tacgacccag ttagccttgt
     2701 gcgggaccat accgtcaagg agaaattggt ggagcggggc atcagtgtgc agtcttacaa
     2761 tggcgacctg ttgtatgaac catgggaaat ttattgtgag aaaggcaagc ctttcactag
     2821 cttcaattct tactggaaaa aatgtctgga catgagtatc gaaagcgtga tgcttccccc
     2881 cccctggcgg ctgatgccaa ttactgccgc cgctgaggca atctgggcat gtagcatcga
     2941 agagttgggt ctggagaatg aggctgagaa gccaagtaac gccctgctga ctcgcgcatg
     3001 gtccccagga tggtcaaacg cagacaaact tctgaacgag ttcattgaaa agcagctgat
     3061 tgactacgct aagaactcca aaaaagtcgt tggaaactca acaagcctct tgagccctta
     3121 cctgcacttt ggcgagatct ctgttcggca cgtgttccag tgcgcccgga tgaagcagat
     3181 aatctgggca agagacaaaa acagtgaggg agaggagtct gccgatttgt tcctgagagg
     3241 catcggactc cgggagtaca gccgctacat ttgttttaac ttccctttca cccacgaaca
     3301 gagcctcttg tctcacctgc gcttcttccc atgggacgca gatgttgata aattcaaggc
     3361 ttggcgacag gggcgcaccg ggtatccact tgtcgacgct ggcatgaggg agctgtgggc
     3421 cacggggtgg atgcataata ggatcagagt gattgtgtct tctttcgccg tcaagttctt
     3481 gttgctccca tggaaatggg gcatgaagta cttctgggac acgctgctgg acgctgacct
     3541 tgagtgcgac attcttgggt ggcagtacat ttccggatct attcctgatg ggcacgaact
     3601 cgataggctc gacaaccccg ctttgcaggg ggctaaatac gacccagaag gggaatacat
     3661 ccgccagtgg ctccccgagc tggcaaggct gcctaccgag tggattcatc atccctggga
     3721 cgcaccgttg actgttttga aggcaagcgg cgtagagttg ggtaccaatt acgcaaagcc
     3781 tatcgtcgat attgacaccg caagggagct gctggcaaag gctatctcta ggacacggga
     3841 ggcccaaatt atgatcggcg ccgcccctga tgagattgta gcagatagct tcgaggcctt
     3901 aggggctaat accattaaag aacctggtct ttgcccatct gtgtcttcta atgaccaaca
     3961 agtaccttcg gctgttggag gaggcgcggg agctagcgga ggtggatctg gtggcggtga
     4021 gggcagagga agtcttctaa catgcggtga cgtggaggag aatcccggcc ctcgtacggg
     4081 cggcagtggt ggcggcagca tgaatggcgc catcggtggc gaccttctcc tgaactttcc
     4141 agacatgagc gttcttgagc gacagagggc tcatctcaaa tacctgaacc cgacgttcga
     4201 tagtcccttg gctggattct ttgctgattc ttctatgatt accggcggcg aaatggattc
     4261 ttatctgagt actgctggtt tgaacctgcc tatgatgtac ggagagacca cagtggaggg
     4321 agatagcaga ctgtccatct ccggaggagg cgcgggaatc gatatggtga gcaagggcga
     4381 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     4441 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     4501 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     4561 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     4621 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     4681 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     4741 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     4801 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     4861 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     4921 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     4981 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     5041 cgccgccggg atcactctcg gcatggacga gctgtataag ggtaaaaaga agaaaaagaa
     5101 gtcaaagaca aagtgtgtaa ttatgtaaga attctgcaga tatccatcac actggcggcc
     5161 gctcgagcat gcatctagaa taatcaacct ctggattaca aaatttgtga aagattgact
     5221 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     5281 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggtta
     5341 gttcttgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     5401 ctgttgggca ctgacaattc cgtggtgttt atttgtgaaa tttgtgatgc tattgcttta
     5461 tttgtaacca tcactagtta ggctttattt gtgaaatttg tgatgctatt gctttatttg
     5521 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     5581 aggttcaggg ggagatgtgg gaggtttttt aaaatcgata aggatcttcc tagagcatgg
     5641 ctacgtagat aagtagcatg gcgggttaat cattaactac aaggaacccc tagtgatgga
     5701 gttggccact ccctctctgc gcgctcgctc gctcactgac tcgctgcgct cggtcgttcg
     5761 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5821 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5881 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5941 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6001 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6061 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6121 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6181 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6241 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6301 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6361 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6421 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6481 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6541 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6601 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6661 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6721 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6781 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6841 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6901 tattaattgt tgccgggaag
//
