LOCUS       sequence-477231-        7858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A recombinant AAV2 plasmid encoding the PiGM-Iq system with CRY2PHR,
            CIBN and EGFP 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 7858)
  TITLE     PiGM-Iq system plasmids
REFERENCE   2  (bases 1 to 7858)
  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-GFAPp-PiGM-Iq(eGFP)
FEATURES             Location/Qualifiers
     source          1..7858
                     /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             join(3953..3955,3956..3958,3959..4669)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3953..3955,3956..3958,3959..4669)
                     /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(3998..4019)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4259..4278)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4606..4627
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4779..4799)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   5282..5411
                     /label=AAV2 ITR
     rep_origin      5593..6048
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5680..5699)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5890..5911
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6074..6178
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6179..6247,6248..7039)
                     /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      7210..7798
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7699..7718
                     /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 gctcacckrg gccgcccggg caaagcccgg gcgkcgggyg rsckttggcw
      241 mgygyagcgg tcacgckscg cktacccacc 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 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     4021 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     4081 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     4141 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     4201 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     4261 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     4321 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     4381 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     4441 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     4501 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     4561 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     4621 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtataagg gtaaaaagaa
     4681 gaaaaagaag tcaaagacaa agtgtgtaat tatgtaatct agaataatca acctctggat
     4741 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     4801 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     4861 tcctccttgt ataaatcctg gttagttctt gccacggcgg aactcatcgc cgcctgcctt
     4921 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gtttatttgt
     4981 gaaatttgtg atgctattgc tttatttgta accatcacta gttaggcttt atttgtgaaa
     5041 tttgtgatgc tattgcttta tttgtaacca tcactagtta ggctttattt gtgaaatttg
     5101 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     5161 ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt aaaatcgata
     5221 aggatcttcc tagagcatgg ctacgtagat aagtagcatg gcgggttaat cattaactac
     5281 aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     5341 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     5401 cgagcgcgca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5461 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc rgtgrgcgrg cgagcgcgca
     5521 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     5581 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     5641 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     5701 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     5761 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     5821 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     5881 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     5941 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     6001 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     6061 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     6121 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     6181 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     6241 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     6301 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     6361 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     6421 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     6481 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     6541 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     6601 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     6661 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     6721 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     6781 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     6841 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     6901 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     6961 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     7021 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     7081 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     7141 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     7201 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     7261 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     7321 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     7381 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     7441 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     7501 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     7561 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     7621 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     7681 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     7741 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     7801 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatg
//
