LOCUS       sequence-461233-        7080 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV encoding the 149 amino acid version PiGM-Ii 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 7080)
  TITLE     PiGM-Iq system plasmids
REFERENCE   2  (bases 1 to 7080)
  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-Ii (149, 535/81, eGFP)
FEATURES             Location/Qualifiers
     source          1..7080
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(2..590)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(82..101)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(761..1552,1553..1621))
                     /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     1384..1403
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1622..1726)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1752..2207)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1889..1910)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2101..2120
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2279..2408
                     /label=AAV2 ITR
     promoter        2490..2938
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             5074..5127
                     /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(5419..5421,5422..5424,5425..6135)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5419..5421,5422..5424,5425..6135)
                     /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(5464..5485)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5725..5744)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6072..6093
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6290..6310)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   6737..6866
                     /label=AAV2 ITR
ORIGIN
        1 atttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
       61 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      121 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      181 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      241 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      301 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      361 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      421 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      481 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      541 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      601 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      661 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      721 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      781 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
      841 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
      901 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
      961 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1021 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1081 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1141 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1201 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1261 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1321 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1381 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1441 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1501 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1561 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1621 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1681 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1741 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1801 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     1861 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     1921 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     1981 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2041 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2101 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2161 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     2221 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2281 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg
     2341 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
     2401 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag
     2461 gaagatctct gcagagggcc ctgcgtatga gtgcaagtgg gttttaggac caggatgagg
     2521 cggggntggg ggtgcctacc tgacgaccga ccccgaccca ctggacaagc acccaacccc
     2581 cattccccaa attgcgcatc ccctatcaga gagggggagg ggaaacagga tgcggcgagg
     2641 cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc ctggcggcgc
     2701 gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca ctcgccggtc ccccgcaaac
     2761 tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc ggcccagccg gaccgcacca
     2821 cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg cgctgcggcg ccggcgactc
     2881 agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg tcgtgcctga gagcgcagtc
     2941 gaattcaagc ttatgttcac ccgcgccgtg agccgactga gcaggaagcg gccgccgtct
     3001 gatatccacg acggcgatgg gagctcaagc agcggccacc agagcctcaa gagcacagcc
     3061 aagtgggcgt cctccctgga gaatcttctg gaagatccag agggcgtgaa gagattcagg
     3121 gaatttctga aaaaggaatt cagtgaggaa aacgtcttgt tttggctagc gtgtgaagat
     3181 ttcaagaaaa cggaggacaa gaagcagatg caggaaaagg ccaagaagat ctacatgacc
     3241 ttcctgtcca ataaggcctc ttcacaagtc aatgtggagg ggcagtctcg gctcactgaa
     3301 aagattctgg aagaaccaca ccctctgatg ttccaaaagc tccaggacca gatcttcaat
     3361 ctcatgaagt atgacagcta cagccgcttc ttgaagtcta ccggtggcgg cagcatgaag
     3421 atggacaaga agacaattgt ctggtttcgc agggacctga gaatcgaaga taaccccgca
     3481 ttggcagcag cagcccatga gggttctgtg ttccctgtct tcatttggtg tccagaggag
     3541 gagggacagt tctaccctgg ccgggccagc agatggtgga tgaagcagtc cttggctcac
     3601 ctgtcacaga gtctcaaggc actgggctcc gatctgacgt tgattaagac ccacaacacg
     3661 attagcgcaa ttctggactg catccgggtg accggcgcaa caaaagtggt tttcaaccac
     3721 ctctacgacc cagttagcct tgtgcgggac cataccgtca aggagaaatt ggtggagcgg
     3781 ggcatcagtg tgcagtctta caatggcgac ctgttgtatg aaccatggga aatttattgt
     3841 gagaaaggca agcctttcac tagcttcaat tcttactgga aaaaatgtct ggacatgagt
     3901 atcgaaagcg tgatgcttcc ccccccctgg cggctgatgc caattactgc cgccgctgag
     3961 gcaatctggg catgtagcat cgaagagttg ggtctggaga atgaggctga gaagccaagt
     4021 aacgccctgc tgactcgcgc atggtcccca ggatggtcaa acgcagacaa acttctgaac
     4081 gagttcattg aaaagcagct gattgactac gctaagaact ccaaaaaagt cgttggaaac
     4141 tcaacaagcc tcttgagccc ttacctgcac tttggcgaga tctctgttcg gcacgtgttc
     4201 cagtgcgccc ggatgaagca gataatctgg gcaagagaca aaaacagtga gggagaggag
     4261 tctgccgatt tgttcctgag aggcatcgga ctccgggagt acagccgcta catttgtttt
     4321 aacttccctt tcacccacga acagagcctc ttgtctcacc tgcgcttctt cccatgggac
     4381 gcagatgttg ataaattcaa ggcttggcga caggggcgca ccgggtatcc acttgtcgac
     4441 gctggcatga gggagctgtg ggccacgggg tggatgcata ataggatcag agtgattgtg
     4501 tcttctttcg ccgtcaagtt cttgttgctc ccatggaaat ggggcatgaa gtacttctgg
     4561 gacacgctgc tggacgctga ccttgagtgc gacattcttg ggtggcagta catttccgga
     4621 tctattcctg atgggcacga actcgatagg ctcgacaacc ccgctttgca gggggctaaa
     4681 tacgacccag aaggggaata catccgccag tggctccccg agctggcaag gctgcctacc
     4741 gagtggattc atcatccctg ggacgcaccg ttgactgttt tgaaggcaag cggcgtagag
     4801 ttgggtacca attacgcaaa gcctatcgtc gatattgaca ccgcaaggga gctgctggca
     4861 aaggctatct ctaggacacg ggaggcccaa attatgatcg gcgccgcccc tgatgagatt
     4921 gtagcagata gcttcgaggc cttaggggct aataccatta aagaacctgg tctttgccca
     4981 tctgtgtctt ctaatgacca acaagtacct tcggctgtcg atcttaagtt ggctggactt
     5041 gatattgcta gcggaggtgg atctggtggc ggtgagggca gaggaagtct tctaacatgc
     5101 ggtgacgtgg aggagaatcc cggccctcgt acgggcggca gtggtggcgg cagcatgaat
     5161 ggcgccatcg gtggcgacct tctcctgaac tttccagaca tgagcgttct tgagcgacag
     5221 agggctcatc tcaaatacct gaacccgacg ttcgatagtc ccttggctgg attctttgct
     5281 gattcttcta tgattaccgg cggcgaaatg gattcttatc tgagtactgc tggtttgaac
     5341 ctgcctatga tgtacggaga gaccacagtg gagggagata gcagactgtc catctccgga
     5401 ggaggcgcgg gaatcgatat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     5461 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     5521 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     5581 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     5641 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     5701 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     5761 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     5821 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     5881 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     5941 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     6001 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     6061 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     6121 gacgagctgt ataagggtaa aaagaagaaa aagaagtcaa agacaaagtg tgtaattatg
     6181 taagaattct gcagatatcc atcacactgg cggccgctcg agcatgcatc tagaataatc
     6241 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     6301 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     6361 ctttcatttt ctcctccttg tataaatcct ggttagttct tgccacggcg gaactcatcg
     6421 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     6481 tgtttatttg tgaaatttgt gatgctattg ctttatttgt aaccatcact agttaggctt
     6541 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     6601 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt
     6661 tttttaaaat cgataaggat cttcctaggg catggctacg tagataagta gcatggcggg
     6721 ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     6781 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     6841 gcctcagtga gcgagcgagc gcgcagctgc attaatgaat cggccaacgc gcggggagag
     6901 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6961 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     7021 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
//
