LOCUS       sequence-477205-        7591 bp    DNA     circular SYN 26-JUN-2025
DEFINITION  A recombinant AAV2 plasmid encoding the light insensitive PiGM-Iq 
            system with CRY2PHR(D387A), CIBN and mCherry as expression marker. 
            hSynapsin promoter for panneuronal expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7591)
  TITLE     PiGM-Iq system plasmids
REFERENCE   2  (bases 1 to 7591)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 26, 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 mCherry)
FEATURES             Location/Qualifiers
     source          1..7591
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   378..507
                     /label=AAV2 ITR
     promoter        589..1036
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             3116..3169
                     /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             3731..4438
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3876..3894)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4144..4163
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(4569..4589)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   5016..5145
                     /label=AAV2 ITR
     rep_origin      5326..5781
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5413..5432)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5623..5644
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5807..5911
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5912..5980,5981..6772)
                     /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(6130..6149)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6943..7531
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7432..7451
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
       61 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      121 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgc
      181 gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg acctttggtc
      241 gcscrgcsgy cacgcksggc ktamccacca cacccgccgc gcttaatgcg ccgctacagg
      301 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
      361 cttcgctatt acgccagctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg
      421 ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag
      481 tggccaactc catcactagg ggttccttgt agttaatgat taacccgcca tgctacttat
      541 ctacgtagcc atgctctagg aagatctctg cagagggccc tgcgtatgag tgcaagtggg
      601 ttttaggacc aggatgaggc ggggtggggg tgcctacctg acgaccgacc ccgacccact
      661 ggacaagcac ccaaccccca ttccccaaat tgcgcatccc ctatcagaga gggggagggg
      721 aaacaggatg cggcgaggcg cgtgcgcact gccagcttca gcaccgcgga cagtgccttc
      781 gcccccgcct ggcggcgcgc gccaccgccg cctcagcact gaaggcgcgc tgacgtcact
      841 cgccggtccc ccgcaaactc cccttcccgg ccaccttggt cgcgtccgcg ccgccgccgg
      901 cccagccgga ccgcaccacg cgaggcgcga gatagggggg cacgggcgcg accatctgcg
      961 ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg ggcagcggag gagtcgtgtc
     1021 gtgcctgaga gcgcagtcga attcaagctt gggatccatg actcctggga agcccaaaac
     1081 tggcaagaaa agcaaacagc aaacttttat caagccttct cctgaggaag cgctgctctg
     1141 ggcagaagca tttgatgaac tgctggccag caaatatggg cttgctgcat tcagagcgtt
     1201 tttaaagtcc gagttctgtg aagaaaatat tgaattctgg ttggcttgcg aagacttcaa
     1261 gaaaaccaaa tcaccacaga aactgtcctc aaaagcaagg aaaatataca ccgacttcat
     1321 cgagaaggaa gctcccaaag agataaacat agactttcaa acaaaaactc tgattgccca
     1381 aaatatccaa gaggctacaa gtggctgctt caccacagcg cagaagaggg tgtacagcct
     1441 gatggagaac aactcttatc ctcgtttctt ggaatcagaa ttttatcagg acttgtgtaa
     1501 aaagccacag atcaccacgg agccccatgc tacaaccggt ggcggcagtg gtggcggcag
     1561 catgaagatg gacaagaaga caattgtctg gtttcgcagg gacctgagaa tcgaagataa
     1621 ccccgcattg gcagcagcag cccatgaggg ttctgtgttc cctgtcttca tttggtgtcc
     1681 agaggaggag ggacagttct accctggccg ggccagcaga tggtggatga agcagtcctt
     1741 ggctcacctg tcacagagtc tcaaggcact gggctccgat ctgacgttga ttaagaccca
     1801 caacacgatt agcgcaattc tggactgcat ccgggtgacc ggcgcaacaa aagtggtttt
     1861 caaccacctc tacgacccag ttagccttgt gcgggaccat accgtcaagg agaaattggt
     1921 ggagcggggc atcagtgtgc agtcttacaa tggcgacctg ttgtatgaac catgggaaat
     1981 ttattgtgag aaaggcaagc ctttcactag cttcaattct tactggaaaa aatgtctgga
     2041 catgagtatc gaaagcgtga tgcttccccc cccctggcgg ctgatgccaa ttactgccgc
     2101 cgctgaggca atctgggcat gtagcatcga agagttgggt ctggagaatg aggctgagaa
     2161 gccaagtaac gccctgctga ctcgcgcatg gtccccagga tggtcaaacg cagacaaact
     2221 tctgaacgag ttcattgaaa agcagctgat tgactacgct aagaactcca aaaaagtcgt
     2281 tggaaactca acaagcctct tgagccctta cctgcacttt ggcgagatct ctgttcggca
     2341 cgtgttccag tgcgcccgga tgaagcagat aatctgggca agagacaaaa acagtgaggg
     2401 agaggagtct gccgatttgt tcctgagagg catcggactc cgggagtaca gccgctacat
     2461 ttgttttaac ttccctttca cccacgaaca gagcctcttg tctcacctgc gcttcttccc
     2521 atgggacgca gatgttgata aattcaaggc ttggcgacag gggcgcaccg ggtatccact
     2581 tgtcgacgct ggcatgaggg agctgtgggc cacggggtgg atgcataata ggatcagagt
     2641 gattgtgtct tctttcgccg tcaagttctt gttgctccca tggaaatggg gcatgaagta
     2701 cttctgggac acgctgctgg ccgctgacct tgagtgcgac attcttgggt ggcagtacat
     2761 ttccggatct attcctgatg ggcacgaact cgataggctc gacaaccccg ctttgcaggg
     2821 ggctaaatac gacccagaag gggaatacat ccgccagtgg ctccccgagc tggcaaggct
     2881 gcctaccgag tggattcatc atccctggga cgcaccgttg actgttttga aggcaagcgg
     2941 cgtagagttg ggtaccaatt acgcaaagcc tatcgtcgat attgacaccg caagggagct
     3001 gctggcaaag gctatctcta ggacacggga ggcccaaatt atgatcggcg ccgcctcagg
     3061 aggaggctca ggaggaggag gcgcgggagc tagcggaggt ggatctggtg gcggtgaggg
     3121 cagaggaagt cttctaacat gcggtgacgt ggaggagaat cccggccctc gtacgggcgg
     3181 cagtggtggc ggcagcatga atggcgccat cggtggcgac cttctcctga actttccaga
     3241 catgagcgtt cttgagcgac agagggctca tctcaaatac ctgaacccga cgttcgatag
     3301 tcccttggct ggattctttg ctgattcttc tatgattacc ggcggcgaaa tggattctta
     3361 tctgagtact gctggtttga acctgcctat gatgtacgga gagaccacag tggagggaga
     3421 tagcagactg tccatctccc cagagaccac gctgggcaca ggaaacttca aagcagccaa
     3481 gtttgataca gagaccaaag actgtaatga agccgctaaa aagatgacca tgaaccggga
     3541 cgatcttgtg gaagagggcg aggaagagaa gtcaaagatt acagaacaga acaatggttc
     3601 caccaagtca atcaaaaaaa tgaagcacaa ggccaaaaaa gaggaaaata atttttccaa
     3661 cgattccagc aaagtgacaa aggaactgga gaagaccgac tacattggag gaggaggcgc
     3721 gggaatcgat atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat
     3781 gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga
     3841 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg
     3901 ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta
     3961 cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa
     4021 gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc
     4081 cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga
     4141 cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc
     4201 cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca
     4261 ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc
     4321 ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga
     4381 acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagga
     4441 tagtgctggt agtgaattcg gtaaaaagaa gaaaaagaag tcaaagacaa agtgtgtaat
     4501 tatgtaatct agaataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     4561 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     4621 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttagttctt
     4681 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     4741 ggcactgaca attccgtggt gtttatttgt gaaatttgtg atgctattgc tttatttgta
     4801 accatcacta gttaggcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     4861 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     4921 agggggagat gtgggaggtt ttttaaaatc gataaggatc ttcctagagc atggctacgt
     4981 agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc
     5041 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     5101 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgca ttaatgaatc
     5161 ggccaacgcg cggggagagg cggtttgcgt attgggmgct cttccgcctc ccgskcwytg
     5221 cccsggcggc ctcagtgagc gagcgagcgc gcagctggcg taatagcgaa gaggcccgca
     5281 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg
     5341 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     5401 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     5461 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     5521 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     5581 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     5641 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     5701 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     5761 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5821 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     5881 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     5941 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     6001 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     6061 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     6121 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     6181 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     6241 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     6301 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     6361 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     6421 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     6481 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     6541 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6601 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6661 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6721 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     6781 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6841 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6901 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6961 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     7021 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     7081 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     7141 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     7201 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     7261 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     7321 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     7381 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     7441 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     7501 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     7561 cctggccttt tgctggcctt ttgctcacat g
//
