LOCUS       sequence-477207-        7867 bp    DNA     circular SYN 08-SEP-2025
DEFINITION  A recombinant AAV2 plasmid encoding the light insensitive PiGM-Iq 
            system with CRY2PHR(D387A), CIBN and mCherry as expression marker. 
            GFAP promoter for astrocyte-selective expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7867)
  TITLE     PiGM-Iq system plasmids
REFERENCE   2  (bases 1 to 7867)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 8, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AAV-GFAPp-PiGM-Iq(D387A mCherry)
FEATURES             Location/Qualifiers
     source          1..7867
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   379..508
                     /label=AAV2 ITR
     CDS             3338..3391
                     /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             3953..4660
                     /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(4098..4116)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4366..4385
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(4791..4811)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   5294..5423
                     /label=AAV2 ITR
     rep_origin      5602..6057
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5689..5708)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5899..5920
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6083..6187
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6188..6256,6257..7048)
                     /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"
     rep_origin      7219..7807
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7708..7727
                     /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 gctcamckrg gccgcccgsg caaagcccss ggcsktsggs grmcyttkgs
      241 tcgcscrgcs gycrcgcksg gskwamcacc acacccgccg cgcttaatgc gccgctacag
      301 ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
      361 tcttcgctat tacgccagct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
      421 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
      481 gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta
      541 tctacgtagc catgctctag gaagatccgg cgcgccgatc taacatatcc tggtgtggag
      601 taggggacgc tgctctgaca gaggctcggg ggcctgagct ggctctgtga gctggggagg
      661 aggcagacag ccaggccttg tctgcaagca gacctggcag cattgggctg gccgcccccc
      721 agggcctcct cttcatgccc agtgaatgac tcaccttggc acagacacaa tgttcggggt
      781 gggcacagtg cctgcttccc gccgcacccc agcccccctc aaatgccttc cgagaagccc
      841 attgagcagg gggcttgcat tgcaccccag cctgacagcc tggcatcttg ggataaaagc
      901 agcacagccc cctaggggct gcccttgctg tgtggcgcca ccggcggtgg agaacaaggc
      961 tctattcagc ctgtgcccag gaaaggggat caggggatgc ccaggcatgg acagtgggtg
     1021 gcaggggggg agaggagggc tgtctgcttc ccagaagtcc aaggacacaa atgggtgagg
     1081 ggagagctct ccccatagct gggctgcggc ccaaccccac cccctcaggc tatgccaggg
     1141 ggtgttgcca ggggcacccg ggcatcgcca gtctagccca ctccttcata aagccctcgc
     1201 atcccaggag cgagcagagc cagagcaggt tggagaggag acgcatcacc tccgctgctc
     1261 gcggggaagc ttgggatcca tgactcctgg gaagcccaaa actggcaaga aaagcaaaca
     1321 gcaaactttt atcaagcctt ctcctgagga agcgctgctc tgggcagaag catttgatga
     1381 actgctggcc agcaaatatg ggcttgctgc attcagagcg tttttaaagt ccgagttctg
     1441 tgaagaaaat attgaattct ggttggcttg cgaagacttc aagaaaacca aatcaccaca
     1501 gaaactgtcc tcaaaagcaa ggaaaatata caccgacttc atcgagaagg aagctcccaa
     1561 agagataaac atagactttc aaacaaaaac tctgattgcc caaaatatcc aagaggctac
     1621 aagtggctgc ttcaccacag cgcagaagag ggtgtacagc ctgatggaga acaactctta
     1681 tcctcgtttc ttggaatcag aattttatca ggacttgtgt aaaaagccac agatcaccac
     1741 ggagccccat gctacaaccg gtggcggcag tggtggcggc agcatgaaga tggacaagaa
     1801 gacaattgtc tggtttcgca gggacctgag aatcgaagat aaccccgcat tggcagcagc
     1861 agcccatgag ggttctgtgt tccctgtctt catttggtgt ccagaggagg agggacagtt
     1921 ctaccctggc cgggccagca gatggtggat gaagcagtcc ttggctcacc tgtcacagag
     1981 tctcaaggca ctgggctccg atctgacgtt gattaagacc cacaacacga ttagcgcaat
     2041 tctggactgc atccgggtga ccggcgcaac aaaagtggtt ttcaaccacc tctacgaccc
     2101 agttagcctt gtgcgggacc ataccgtcaa ggagaaattg gtggagcggg gcatcagtgt
     2161 gcagtcttac aatggcgacc tgttgtatga accatgggaa atttattgtg agaaaggcaa
     2221 gcctttcact agcttcaatt cttactggaa aaaatgtctg gacatgagta tcgaaagcgt
     2281 gatgcttccc cccccctggc ggctgatgcc aattactgcc gccgctgagg caatctgggc
     2341 atgtagcatc gaagagttgg gtctggagaa tgaggctgag aagccaagta acgccctgct
     2401 gactcgcgca tggtccccag gatggtcaaa cgcagacaaa cttctgaacg agttcattga
     2461 aaagcagctg attgactacg ctaagaactc caaaaaagtc gttggaaact caacaagcct
     2521 cttgagccct tacctgcact ttggcgagat ctctgttcgg cacgtgttcc agtgcgcccg
     2581 gatgaagcag ataatctggg caagagacaa aaacagtgag ggagaggagt ctgccgattt
     2641 gttcctgaga ggcatcggac tccgggagta cagccgctac atttgtttta acttcccttt
     2701 cacccacgaa cagagcctct tgtctcacct gcgcttcttc ccatgggacg cagatgttga
     2761 taaattcaag gcttggcgac aggggcgcac cgggtatcca cttgtcgacg ctggcatgag
     2821 ggagctgtgg gccacggggt ggatgcataa taggatcaga gtgattgtgt cttctttcgc
     2881 cgtcaagttc ttgttgctcc catggaaatg gggcatgaag tacttctggg acacgctgct
     2941 ggacgctgac cttgagtgcg acattcttgg gtggcagtac atttccggat ctattcctga
     3001 tgggcacgaa ctcgataggc tcgacaaccc cgctttgcag ggggctaaat acgacccaga
     3061 aggggaatac atccgccagt ggctccccga gctggcaagg ctgcctaccg agtggattca
     3121 tcatccctgg gacgcaccgt tgactgtttt gaaggcaagc ggcgtagagt tgggtaccaa
     3181 ttacgcaaag cctatcgtcg atattgacac cgcaagggag ctgctggcaa aggctatctc
     3241 taggacacgg gaggcccaaa ttatgatcgg cgccgcctca ggaggaggct caggaggagg
     3301 aggcgcggga gctagcggag gtggatctgg tggcggtgag ggcagaggaa gtcttctaac
     3361 atgcggtgac gtggaggaga atcccggccc tcgtacgggc ggcagtggtg gcggcagcat
     3421 gaatggcgcc atcggtggcg accttctcct gaactttcca gacatgagcg ttcttgagcg
     3481 acagagggct catctcaaat acctgaaccc gacgttcgat agtcccttgg ctggattctt
     3541 tgctgattct tctatgatta ccggcggcga aatggattct tatctgagta ctgctggttt
     3601 gaacctgcct atgatgtacg gagagaccac agtggaggga gatagcagac tgtccatctc
     3661 cccagagacc acgctgggca caggaaactt caaagcagcc aagtttgata cagagaccaa
     3721 agactgtaat gaagccgcta aaaagatgac catgaaccgg gacgatcttg tggaagaggg
     3781 cgaggaagag aagtcaaaga ttacagaaca gaacaatggt tccaccaagt caatcaaaaa
     3841 aatgaagcac aaggccaaaa aagaggaaaa taatttttcc aacgattcca gcaaagtgac
     3901 aaaggaactg gagaagaccg actacattgg aggaggaggc gcgggaatcg atatggtgag
     3961 caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat
     4021 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     4081 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg
     4141 ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga
     4201 catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     4261 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt
     4321 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa
     4381 gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa
     4441 gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa
     4501 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa
     4561 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga
     4621 gggccgccac tccaccggcg gcatggacga gctgtacaag gatagtgctg gtagtgaatt
     4681 cggtaaaaag aagaaaaaga agtcaaagac aaagtgtgta attatgtaat ctagaataat
     4741 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     4801 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     4861 gctttcattt tctcctcctt gtataaatcc tggttagttc ttgccacggc ggaactcatc
     4921 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     4981 gtgtttattt gtgaaatttg tgatgctatt gctttatttg taaccatcac tagttaggct
     5041 ttatttgtga aatttgtgat gctattgctt tatttgtaac catcactagt taggctttat
     5101 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     5161 taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt
     5221 ttaaaatcga taaggatctt cctagagcat ggctacgtag ataagtagca tggcgggtta
     5281 atcattaact acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     5341 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     5401 tcagtgagcg agcgagcgcg cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5461 gyttgcgwmy trgggkcgcc csmcgcccgg gctttgcccg ggcggcctca gtgagcgagc
     5521 gagcgcgcag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     5581 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     5641 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     5701 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     5761 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     5821 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     5881 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     5941 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     6001 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     6061 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     6121 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     6181 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     6241 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     6301 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     6361 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     6421 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     6481 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     6541 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     6601 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     6661 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     6721 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     6781 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     6841 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     6901 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6961 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     7021 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     7081 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     7141 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     7201 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     7261 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     7321 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     7381 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     7441 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     7501 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     7561 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     7621 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     7681 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     7741 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     7801 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     7861 tcacatg
//
