LOCUS       sequence-441622-        7269 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A recombinant AAV2 plasmid encoding the light insensitive PiGM-Iq 
            system with CRY2PHR(D387A), CIBN and EGFP as expression marker. 
            hSynapsin promoter for panneuronal expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7269)
  TITLE     PiGM-Iq system plasmids
REFERENCE   2  (bases 1 to 7269)
  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(D387A eGFP)
FEATURES             Location/Qualifiers
     source          1..7269
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     316..336
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(446..1156,1157..1159,1160..1162))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(446..1156,1157..1159,1160..1162))
                     /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(488..509)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     837..856
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1096..1117
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(1724..1777)
                     /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"
     promoter        complement(3857..4304)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   4386..4515
                     /label=AAV2 ITR
     rep_origin      4587..5042
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4674..4693)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4884..4905
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5068..5172
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5173..5241,5242..6033)
                     /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(5391..5410)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6204..6792
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6693..6712
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   7029..7158
                     /label=AAV2 ITR
ORIGIN
        1 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
       61 aatagcatca caaatttcac aaataaagcc taactagtga tggttacaaa taaagcaata
      121 gcatcacaaa tttcacaaat aaacaccacg gaattgtcag tgcccaacag ccgagcccct
      181 gtccagcagc gggcaaggca ggcggcgatg agttccgccg tggcaagaac taaccaggat
      241 ttatacaagg aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt
      301 aaagcagcgt atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt
      361 tcacaaattt tgtaatccag aggttgatta ttctagatta cataattaca cactttgtct
      421 ttgacttctt tttcttcttt ttacccttat acagctcgtc catgccgaga gtgatcccgg
      481 cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg tctttgctca
      541 gggcggactg ggtgctcagg tagtggttgt cgggcagcag cacggggccg tcgccgatgg
      601 gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc gtcctcgatg ttgtggcgga
      661 tcttgaagtt caccttgatg ccgttcttct gcttgtcggc catgatatag acgttgtggc
      721 tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc
      781 ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg gcgcgggtct
      841 tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg
      901 cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcggctgaag cactgcacgc
      961 cgtaggtcag ggtggtcacg agggtgggcc agggcacggg cagcttgccg gtggtgcaga
     1021 tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg gacacgctga
     1081 acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc ccggtgaaca
     1141 gctcctcgcc cttgctcacc atatcgattc ccgcgcctcc tcctccaatg tagtcggtct
     1201 tctccagttc ctttgtcact ttgctggaat cgttggaaaa attattttcc tcttttttgg
     1261 ccttgtgctt catttttttg attgacttgg tggaaccatt gttctgttct gtaatctttg
     1321 acttctcttc ctcgccctct tccacaagat cgtcccggtt catggtcatc tttttagcgg
     1381 cttcattaca gtctttggtc tctgtatcaa acttggctgc tttgaagttt cctgtgccca
     1441 gcgtggtctc tggggagatg gacagtctgc tatctccctc cactgtggtc tctccgtaca
     1501 tcataggcag gttcaaacca gcagtactca gataagaatc catttcgccg ccggtaatca
     1561 tagaagaatc agcaaagaat ccagccaagg gactatcgaa cgtcgggttc aggtatttga
     1621 gatgagccct ctgtcgctca agaacgctca tgtctggaaa gttcaggaga aggtcgccac
     1681 cgatggcgcc attcatgctg ccgccaccac tgccgcccgt acgagggccg ggattctcct
     1741 ccacgtcacc gcatgttaga agacttcctc tgccctcacc gccaccagat ccacctccgc
     1801 tagctcccgc gcctcctcct cctgagcctc ctcctgaggc ggcgccgatc ataatttggg
     1861 cctcccgtgt cctagagata gcctttgcca gcagctccct tgcggtgtca atatcgacga
     1921 taggctttgc gtaattggta cccaactcta cgccgcttgc cttcaaaaca gtcaacggtg
     1981 cgtcccaggg atgatgaatc cactcggtag gcagccttgc cagctcgggg agccactggc
     2041 ggatgtattc cccttctggg tcgtatttag ccccctgcaa agcggggttg tcgagcctat
     2101 cgagttcgtg cccatcagga atagatccgg aaatgtactg ccacccaaga atgtcgcact
     2161 caaggtcagc ggccagcagc gtgtcccaga agtacttcat gccccatttc catgggagca
     2221 acaagaactt gacggcgaaa gaagacacaa tcactctgat cctattatgc atccaccccg
     2281 tggcccacag ctccctcatg ccagcgtcga caagtggata cccggtgcgc ccctgtcgcc
     2341 aagccttgaa tttatcaaca tctgcgtccc atgggaagaa gcgcaggtga gacaagaggc
     2401 tctgttcgtg ggtgaaaggg aagttaaaac aaatgtagcg gctgtactcc cggagtccga
     2461 tgcctctcag gaacaaatcg gcagactcct ctccctcact gtttttgtct cttgcccaga
     2521 ttatctgctt catccgggcg cactggaaca cgtgccgaac agagatctcg ccaaagtgca
     2581 ggtaagggct caagaggctt gttgagtttc caacgacttt tttggagttc ttagcgtagt
     2641 caatcagctg cttttcaatg aactcgttca gaagtttgtc tgcgtttgac catcctgggg
     2701 accatgcgcg agtcagcagg gcgttacttg gcttctcagc ctcattctcc agacccaact
     2761 cttcgatgct acatgcccag attgcctcag cggcggcagt aattggcatc agccgccagg
     2821 gggggggaag catcacgctt tcgatactca tgtccagaca ttttttccag taagaattga
     2881 agctagtgaa aggcttgcct ttctcacaat aaatttccca tggttcatac aacaggtcgc
     2941 cattgtaaga ctgcacactg atgccccgct ccaccaattt ctccttgacg gtatggtccc
     3001 gcacaaggct aactgggtcg tagaggtggt tgaaaaccac ttttgttgcg ccggtcaccc
     3061 ggatgcagtc cagaattgcg ctaatcgtgt tgtgggtctt aatcaacgtc agatcggagc
     3121 ccagtgcctt gagactctgt gacaggtgag ccaaggactg cttcatccac catctgctgg
     3181 cccggccagg gtagaactgt ccctcctcct ctggacacca aatgaagaca gggaacacag
     3241 aaccctcatg ggctgctgct gccaatgcgg ggttatcttc gattctcagg tccctgcgaa
     3301 accagacaat tgtcttcttg tccatcttca tgctgccgcc accactgccg ccaccggttg
     3361 tagcatgggg ctccgtggtg atctgtggct ttttacacaa gtcctgataa aattctgatt
     3421 ccaagaaacg aggataagag ttgttctcca tcaggctgta caccctcttc tgcgctgtgg
     3481 tgaagcagcc acttgtagcc tcttggatat tttgggcaat cagagttttt gtttgaaagt
     3541 ctatgtttat ctctttggga gcttccttct cgatgaagtc ggtgtatatt ttccttgctt
     3601 ttgaggacag tttctgtggt gatttggttt tcttgaagtc ttcgcaagcc aaccagaatt
     3661 caatattttc ttcacagaac tcggacttta aaaacgctct gaatgcagca agcccatatt
     3721 tgctggccag cagttcatca aatgcttctg cccagagcag cgcttcctca ggagaaggct
     3781 tgataaaagt ttgctgtttg cttttcttgc cagttttggg cttcccagga gtcatggatc
     3841 ccaagcttga attcgactgc gctctcaggc acgacacgac tcctccgctg cccaccgcag
     3901 actgaggcag cgctgagtcg ccggcgccgc agcgcagatg gtcgcgcccg tgccccccta
     3961 tctcgcgcct cgcgtggtgc ggtccggctg ggccggcggc ggcgcggacg cgaccaaggt
     4021 ggccgggaag gggagtttgc gggggaccgg cgagtgacgt cagcgcgcct tcagtgctga
     4081 ggcggcggtg gcgcgcgccg ccaggcgggg gcgaaggcac tgtccgcggt gctgaagctg
     4141 gcagtgcgca cgcgcctcgc cgcatcctgt ttcccctccc cctctctgat aggggatgcg
     4201 caatttgggg aatgggggtt gggtgcttgt ccagtgggtc ggggtcggtc gtcaggtagg
     4261 cacccccacc ccgcctcatc ctggtcctaa aacccacttg cactcatacg cagggccctc
     4321 tgcagagatc ttcctagagc atggctacgt agataagtag catggcgggt taatcattaa
     4381 ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     4441 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     4501 cgagcgagcg cgcagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     4561 gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
     4621 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4681 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4741 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4801 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4861 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4921 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     4981 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     5041 ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5101 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5161 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5221 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5281 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5341 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5401 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5461 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5521 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5581 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5641 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5701 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5761 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5821 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5881 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5941 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6001 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6061 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6121 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6181 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6241 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6301 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6361 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6421 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6481 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6541 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6601 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6661 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6721 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6781 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6841 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6901 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     6961 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     7021 taatgcagct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg
     7081 cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact
     7141 ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc
     7201 catgctctag gaagatcctt atcgatttta aaaaacctcc cacatctccc cctgaacctg
     7261 aaacataaa
//
