LOCUS       sequence-477209-        7869 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A recombinant AAV2 plasmid encoding the PiGM-Iq system with CRY2PHR,
            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 7869)
  TITLE     PiGM-Iq system plasmids
REFERENCE   2  (bases 1 to 7869)
  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(mCherry)
FEATURES             Location/Qualifiers
     source          1..7869
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   378..507
                     /label=AAV2 ITR
     CDS             3337..3390
                     /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             3952..4659
                     /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(4097..4115)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4365..4384
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(4790..4810)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   5293..5422
                     /label=AAV2 ITR
     rep_origin      5604..6059
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5691..5710)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5901..5922
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6085..6189
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6190..6258,6259..7050)
                     /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      7221..7809
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7710..7729
                     /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 gctcacctgg gscgcccggg caaagcccss ngcrwtsggs gksckttrgs
      241 kygcmgsgky crcgctgcgc gtaaccacca 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 aagatccggc gcgccgatct aacatatcct ggtgtggagt
      601 aggggacgct gctctgacag aggctcgggg gcctgagctg gctctgtgag ctggggagga
      661 ggcagacagc caggccttgt ctgcaagcag acctggcagc attgggctgg ccgcccccca
      721 gggcctcctc ttcatgccca gtgaatgact caccttggca cagacacaat gttcggggtg
      781 ggcacagtgc ctgcttcccg ccgcacccca gcccccctca aatgccttcc gagaagccca
      841 ttgagcaggg ggcttgcatt gcaccccagc ctgacagcct ggcatcttgg gataaaagca
      901 gcacagcccc ctaggggctg cccttgctgt gtggcgccac cggcggtgga gaacaaggct
      961 ctattcagcc tgtgcccagg aaaggggatc aggggatgcc caggcatgga cagtgggtgg
     1021 caggggggga gaggagggct gtctgcttcc cagaagtcca aggacacaaa tgggtgaggg
     1081 gagagctctc cccatagctg ggctgcggcc caaccccacc ccctcaggct atgccagggg
     1141 gtgttgccag gggcacccgg gcatcgccag tctagcccac tccttcataa agccctcgca
     1201 tcccaggagc gagcagagcc agagcaggtt ggagaggaga cgcatcacct ccgctgctcg
     1261 cggggaagct tgggatccat gactcctggg aagcccaaaa ctggcaagaa aagcaaacag
     1321 caaactttta tcaagccttc tcctgaggaa gcgctgctct gggcagaagc atttgatgaa
     1381 ctgctggcca gcaaatatgg gcttgctgca ttcagagcgt ttttaaagtc cgagttctgt
     1441 gaagaaaata ttgaattctg gttggcttgc gaagacttca agaaaaccaa atcaccacag
     1501 aaactgtcct caaaagcaag gaaaatatac accgacttca tcgagaagga agctcccaaa
     1561 gagataaaca tagactttca aacaaaaact ctgattgccc aaaatatcca agaggctaca
     1621 agtggctgct tcaccacagc gcagaagagg gtgtacagcc tgatggagaa caactcttat
     1681 cctcgtttct tggaatcaga attttatcag gacttgtgta aaaagccaca gatcaccacg
     1741 gagccccatg ctacaaccgg tggcggcagt ggtggcggca gcatgaagat ggacaagaag
     1801 acaattgtct ggtttcgcag ggacctgaga atcgaagata accccgcatt ggcagcagca
     1861 gcccatgagg gttctgtgtt ccctgtcttc atttggtgtc cagaggagga gggacagttc
     1921 taccctggcc gggccagcag atggtggatg aagcagtcct tggctcacct gtcacagagt
     1981 ctcaaggcac tgggctccga tctgacgttg attaagaccc acaacacgat tagcgcaatt
     2041 ctggactgca tccgggtgac cggcgcaaca aaagtggttt tcaaccacct ctacgaccca
     2101 gttagccttg tgcgggacca taccgtcaag gagaaattgg tggagcgggg catcagtgtg
     2161 cagtcttaca atggcgacct gttgtatgaa ccatgggaaa tttattgtga gaaaggcaag
     2221 cctttcacta gcttcaattc ttactggaaa aaatgtctgg acatgagtat cgaaagcgtg
     2281 atgcttcccc ccccctggcg gctgatgcca attactgccg ccgctgaggc aatctgggca
     2341 tgtagcatcg aagagttggg tctggagaat gaggctgaga agccaagtaa cgccctgctg
     2401 actcgcgcat ggtccccagg atggtcaaac gcagacaaac ttctgaacga gttcattgaa
     2461 aagcagctga ttgactacgc taagaactcc aaaaaagtcg ttggaaactc aacaagcctc
     2521 ttgagccctt acctgcactt tggcgagatc tctgttcggc acgtgttcca gtgcgcccgg
     2581 atgaagcaga taatctgggc aagagacaaa aacagtgagg gagaggagtc tgccgatttg
     2641 ttcctgagag gcatcggact ccgggagtac agccgctaca tttgttttaa cttccctttc
     2701 acccacgaac agagcctctt gtctcacctg cgcttcttcc catgggacgc agatgttgat
     2761 aaattcaagg cttggcgaca ggggcgcacc gggtatccac ttgtcgacgc tggcatgagg
     2821 gagctgtggg ccacggggtg gatgcataat aggatcagag tgattgtgtc ttctttcgcc
     2881 gtcaagttct tgttgctccc atggaaatgg ggcatgaagt acttctggga cacgctgctg
     2941 gccgctgacc ttgagtgcga cattcttggg tggcagtaca tttccggatc tattcctgat
     3001 gggcacgaac tcgataggct cgacaacccc gctttgcagg gggctaaata cgacccagaa
     3061 ggggaataca tccgccagtg gctccccgag ctggcaaggc tgcctaccga gtggattcat
     3121 catccctggg acgcaccgtt gactgttttg aaggcaagcg gcgtagagtt gggtaccaat
     3181 tacgcaaagc ctatcgtcga tattgacacc gcaagggagc tgctggcaaa ggctatctct
     3241 aggacacggg aggcccaaat tatgatcggc gccgcctcag gaggaggctc aggaggagga
     3301 ggcgcgggag ctagcggagg tggatctggt ggcggtgagg gcagaggaag tcttctaaca
     3361 tgcggtgacg tggaggagaa tcccggccct cgtacgggcg gcagtggtgg cggcagcatg
     3421 aatggcgcca tcggtggcga ccttctcctg aactttccag acatgagcgt tcttgagcga
     3481 cagagggctc atctcaaata cctgaacccg acgttcgata gtcccttggc tggattcttt
     3541 gctgattctt ctatgattac cggcggcgaa atggattctt atctgagtac tgctggtttg
     3601 aacctgccta tgatgtacgg agagaccaca gtggagggag atagcagact gtccatctcc
     3661 ccagagacca cgctgggcac aggaaacttc aaagcagcca agtttgatac agagaccaaa
     3721 gactgtaatg aagccgctaa aaagatgacc atgaaccggg acgatcttgt ggaagagggc
     3781 gaggaagaga agtcaaagat tacagaacag aacaatggtt ccaccaagtc aatcaaaaaa
     3841 atgaagcaca aggccaaaaa agaggaaaat aatttttcca acgattccag caaagtgaca
     3901 aaggaactgg agaagaccga ctacattgga ggaggaggcg cgggaatcga tatggtgagc
     3961 aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg
     4021 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     4081 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg
     4141 gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac
     4201 atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     4261 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc
     4321 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag
     4381 aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag
     4441 ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag
     4501 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag
     4561 ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag
     4621 ggccgccact ccaccggcgg catggacgag ctgtacaagg atagtgctgg tagtgaattc
     4681 ggtaaaaaga agaaaaagaa gtcaaagaca aagtgtgtaa ttatgtaatc tagaataatc
     4741 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     4801 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     4861 ctttcatttt ctcctccttg tataaatcct ggttagttct tgccacggcg gaactcatcg
     4921 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     4981 tgtttatttg tgaaatttgt gatgctattg ctttatttgt aaccatcact agttaggctt
     5041 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc atcactagtt aggctttatt
     5101 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     5161 aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt
     5221 taaaatcgat aaggatcttc ctagagcatg gctacgtaga taagtagcat ggcgggttaa
     5281 tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     5341 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     5401 cagtgagcga gcgagcgcgc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5461 tttgcgtatt gggcgctctt ccgcttcctc gctcwctgmc csggcggcct crgtgagcga
     5521 gcgagcgcgc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     5581 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5641 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     5701 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     5761 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     5821 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     5881 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     5941 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     6001 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     6061 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     6121 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     6181 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6241 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6301 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     6361 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     6421 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     6481 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6541 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6601 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6661 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6721 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6781 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6841 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6901 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6961 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     7021 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     7081 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     7141 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7201 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7261 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7321 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     7381 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7441 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7501 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7561 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7621 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7681 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7741 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7801 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7861 gctcacatg
//
