LOCUS       sequence-477210-        7855 bp    DNA     circular SYN 26-JUN-2025
DEFINITION  A recombinant AAV2 plasmid encoding the light insensitive PiGM-Iq 
            system with CRY2PHR(D387A), 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 7855)
  TITLE     PiGM-Iq system plasmids
REFERENCE   2  (bases 1 to 7855)
  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-GFAPp-PiGM-Iq(D387A eGFP)
FEATURES             Location/Qualifiers
     source          1..7855
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   376..505
                     /label=AAV2 ITR
     CDS             3335..3388
                     /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(3950..3952,3953..3955,3956..4666)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3950..3952,3953..3955,3956..4666)
                     /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(3995..4016)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4256..4275)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4603..4624
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4776..4796)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   5279..5408
                     /label=AAV2 ITR
     rep_origin      5590..6045
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5677..5696)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5887..5908
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6071..6175
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6176..6244,6245..7036)
                     /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      7207..7795
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7696..7715
                     /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 gcccsgsyym cgckgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
      301 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
      361 tcgctattac gccagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg
      421 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
      481 gccaactcca tcactagggg ttccttgtag ttaatgatta acccgccatg ctacttatct
      541 acgtagccat gctctaggaa gatccggcgc gccgatctaa catatcctgg tgtggagtag
      601 gggacgctgc tctgacagag gctcgggggc ctgagctggc tctgtgagct ggggaggagg
      661 cagacagcca ggccttgtct gcaagcagac ctggcagcat tgggctggcc gccccccagg
      721 gcctcctctt catgcccagt gaatgactca ccttggcaca gacacaatgt tcggggtggg
      781 cacagtgcct gcttcccgcc gcaccccagc ccccctcaaa tgccttccga gaagcccatt
      841 gagcaggggg cttgcattgc accccagcct gacagcctgg catcttggga taaaagcagc
      901 acagccccct aggggctgcc cttgctgtgt ggcgccaccg gcggtggaga acaaggctct
      961 attcagcctg tgcccaggaa aggggatcag gggatgccca ggcatggaca gtgggtggca
     1021 gggggggaga ggagggctgt ctgcttccca gaagtccaag gacacaaatg ggtgagggga
     1081 gagctctccc catagctggg ctgcggccca accccacccc ctcaggctat gccagggggt
     1141 gttgccaggg gcacccgggc atcgccagtc tagcccactc cttcataaag ccctcgcatc
     1201 ccaggagcga gcagagccag agcaggttgg agaggagacg catcacctcc gctgctcgcg
     1261 gggaagcttg ggatccatga ctcctgggaa gcccaaaact ggcaagaaaa gcaaacagca
     1321 aacttttatc aagccttctc ctgaggaagc gctgctctgg gcagaagcat ttgatgaact
     1381 gctggccagc aaatatgggc ttgctgcatt cagagcgttt ttaaagtccg agttctgtga
     1441 agaaaatatt gaattctggt tggcttgcga agacttcaag aaaaccaaat caccacagaa
     1501 actgtcctca aaagcaagga aaatatacac cgacttcatc gagaaggaag ctcccaaaga
     1561 gataaacata gactttcaaa caaaaactct gattgcccaa aatatccaag aggctacaag
     1621 tggctgcttc accacagcgc agaagagggt gtacagcctg atggagaaca actcttatcc
     1681 tcgtttcttg gaatcagaat tttatcagga cttgtgtaaa aagccacaga tcaccacgga
     1741 gccccatgct acaaccggtg gcggcagtgg tggcggcagc atgaagatgg acaagaagac
     1801 aattgtctgg tttcgcaggg acctgagaat cgaagataac cccgcattgg cagcagcagc
     1861 ccatgagggt tctgtgttcc ctgtcttcat ttggtgtcca gaggaggagg gacagttcta
     1921 ccctggccgg gccagcagat ggtggatgaa gcagtccttg gctcacctgt cacagagtct
     1981 caaggcactg ggctccgatc tgacgttgat taagacccac aacacgatta gcgcaattct
     2041 ggactgcatc cgggtgaccg gcgcaacaaa agtggttttc aaccacctct acgacccagt
     2101 tagccttgtg cgggaccata ccgtcaagga gaaattggtg gagcggggca tcagtgtgca
     2161 gtcttacaat ggcgacctgt tgtatgaacc atgggaaatt tattgtgaga aaggcaagcc
     2221 tttcactagc ttcaattctt actggaaaaa atgtctggac atgagtatcg aaagcgtgat
     2281 gcttcccccc ccctggcggc tgatgccaat tactgccgcc gctgaggcaa tctgggcatg
     2341 tagcatcgaa gagttgggtc tggagaatga ggctgagaag ccaagtaacg ccctgctgac
     2401 tcgcgcatgg tccccaggat ggtcaaacgc agacaaactt ctgaacgagt tcattgaaaa
     2461 gcagctgatt gactacgcta agaactccaa aaaagtcgtt ggaaactcaa caagcctctt
     2521 gagcccttac ctgcactttg gcgagatctc tgttcggcac gtgttccagt gcgcccggat
     2581 gaagcagata atctgggcaa gagacaaaaa cagtgaggga gaggagtctg ccgatttgtt
     2641 cctgagaggc atcggactcc gggagtacag ccgctacatt tgttttaact tccctttcac
     2701 ccacgaacag agcctcttgt ctcacctgcg cttcttccca tgggacgcag atgttgataa
     2761 attcaaggct tggcgacagg ggcgcaccgg gtatccactt gtcgacgctg gcatgaggga
     2821 gctgtgggcc acggggtgga tgcataatag gatcagagtg attgtgtctt ctttcgccgt
     2881 caagttcttg ttgctcccat ggaaatgggg catgaagtac ttctgggaca cgctgctggc
     2941 cgctgacctt gagtgcgaca ttcttgggtg gcagtacatt tccggatcta ttcctgatgg
     3001 gcacgaactc gataggctcg acaaccccgc tttgcagggg gctaaatacg acccagaagg
     3061 ggaatacatc cgccagtggc tccccgagct ggcaaggctg cctaccgagt ggattcatca
     3121 tccctgggac gcaccgttga ctgttttgaa ggcaagcggc gtagagttgg gtaccaatta
     3181 cgcaaagcct atcgtcgata ttgacaccgc aagggagctg ctggcaaagg ctatctctag
     3241 gacacgggag gcccaaatta tgatcggcgc cgcctcagga ggaggctcag gaggaggagg
     3301 cgcgggagct agcggaggtg gatctggtgg cggtgagggc agaggaagtc ttctaacatg
     3361 cggtgacgtg gaggagaatc ccggccctcg tacgggcggc agtggtggcg gcagcatgaa
     3421 tggcgccatc ggtggcgacc ttctcctgaa ctttccagac atgagcgttc ttgagcgaca
     3481 gagggctcat ctcaaatacc tgaacccgac gttcgatagt cccttggctg gattctttgc
     3541 tgattcttct atgattaccg gcggcgaaat ggattcttat ctgagtactg ctggtttgaa
     3601 cctgcctatg atgtacggag agaccacagt ggagggagat agcagactgt ccatctcccc
     3661 agagaccacg ctgggcacag gaaacttcaa agcagccaag tttgatacag agaccaaaga
     3721 ctgtaatgaa gccgctaaaa agatgaccat gaaccgggac gatcttgtgg aagagggcga
     3781 ggaagagaag tcaaagatta cagaacagaa caatggttcc accaagtcaa tcaaaaaaat
     3841 gaagcacaag gccaaaaaag aggaaaataa tttttccaac gattccagca aagtgacaaa
     3901 ggaactggag aagaccgact acattggagg aggaggcgcg ggaatcgata tggtgagcaa
     3961 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     4021 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     4081 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     4141 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     4201 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     4261 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     4321 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     4381 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     4441 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     4501 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     4561 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     4621 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tataagggta aaaagaagaa
     4681 aaagaagtca aagacaaagt gtgtaattat gtaatctaga ataatcaacc tctggattac
     4741 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     4801 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     4861 tccttgtata aatcctggtt agttcttgcc acggcggaac tcatcgccgc ctgccttgcc
     4921 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt tatttgtgaa
     4981 atttgtgatg ctattgcttt atttgtaacc atcactagtt aggctttatt tgtgaaattt
     5041 gtgatgctat tgctttattt gtaaccatca ctagttaggc tttatttgtg aaatttgtga
     5101 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     5161 cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa atcgataagg
     5221 atcttcctag agcatggcta cgtagataag tagcatggcg ggttaatcat taactacaag
     5281 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     5341 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     5401 gcgcgcagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5461 gctcttccgc ctcctcgskc wytgmccsgg cggcctcagt gagcgagcga gcgcgcagct
     5521 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     5581 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     5641 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     5701 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     5761 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     5821 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     5881 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     5941 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     6001 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     6061 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     6121 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     6181 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     6241 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     6301 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     6361 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     6421 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     6481 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     6541 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     6601 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     6661 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     6721 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     6781 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6841 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6901 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6961 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     7021 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     7081 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     7141 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     7201 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     7261 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     7321 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     7381 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     7441 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     7501 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     7561 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     7621 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     7681 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     7741 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7801 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatg
//
