LOCUS       sequence-441742-        7277 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. hSynapsin promoter for 
            panneuronal expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7277)
  TITLE     PiGM-Iq system plasmids
REFERENCE   2  (bases 1 to 7277)
  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-hSyn-PiGM-Iq(eGFP)
FEATURES             Location/Qualifiers
     source          1..7277
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      72..527
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(159..178)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     369..390
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        553..657
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(658..726,727..1518)
                     /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(876..895)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1689..2277
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2178..2197
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2514..2643
                     /label=AAV2 ITR
     primer_bind     3070..3090
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3200..3910,3911..3913,3914..3916))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(3200..3910,3911..3913,3914..3916))
                     /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(3242..3263)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3591..3610
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3850..3871
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(4478..4531)
                     /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"
     promoter        complement(6611..7058)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   7140..7270
                     /label=AAV2 ITR
ORIGIN
        1 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
       61 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
      121 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      181 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      241 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      301 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      361 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      421 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      481 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
      541 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
      601 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
      661 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
      721 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
      781 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
      841 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
      901 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
      961 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     1021 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     1081 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     1141 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     1201 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     1261 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     1321 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     1381 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     1441 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     1501 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     1561 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     1621 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     1681 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     1741 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1801 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     1861 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     1921 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     1981 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     2041 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2101 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     2161 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     2221 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     2281 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     2341 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     2401 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     2461 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctgcgcg
     2521 ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc
     2581 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
     2641 ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc tctaggaaga
     2701 tccttatcga ttttaaaaaa cctcccacat ctccccctga acctgaaaca taaaatgaat
     2761 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     2821 atcacaaatt tcacaaataa agcctaacta gtgatggtta caaataaagc aatagcatca
     2881 caaatttcac aaataaacac cacggaattg tcagtgccca acagccgagc ccctgtccag
     2941 cagcgggcaa ggcaggcggc gatgagttcc gccgtggcaa gaactaacca ggatttatac
     3001 aaggaggaga aaatgaaagc catacgggaa gcaatagcat gatacaaagg cattaaagca
     3061 gcgtatccac atagcgtaaa aggagcaaca tagttaagaa taccagtcaa tctttcacaa
     3121 attttgtaat ccagaggttg attattctag attacataat tacacacttt gtctttgact
     3181 tctttttctt ctttttaccc ttatacagct cgtccatgcc gagagtgatc ccggcggcgg
     3241 tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg
     3301 actgggtgct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt
     3361 tctgctggta gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg cggatcttga
     3421 agttcacctt gatgccgttc ttctgcttgt cggccatgat atagacgttg tggctgttgt
     3481 agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca
     3541 gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt
     3601 tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact
     3661 tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc acgccgtagg
     3721 tcagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatgaact
     3781 tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt
     3841 ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct
     3901 cgcccttgct caccatatcg attcccgcgc ctcctcctcc aatgtagtcg gtcttctcca
     3961 gttcctttgt cactttgctg gaatcgttgg aaaaattatt ttcctctttt ttggccttgt
     4021 gcttcatttt tttgattgac ttggtggaac cattgttctg ttctgtaatc tttgacttct
     4081 cttcctcgcc ctcttccaca agatcgtccc ggttcatggt catcttttta gcggcttcat
     4141 tacagtcttt ggtctctgta tcaaacttgg ctgctttgaa gtttcctgtg cccagcgtgg
     4201 tctctgggga gatggacagt ctgctatctc cctccactgt ggtctctccg tacatcatag
     4261 gcaggttcaa accagcagta ctcagataag aatccatttc gccgccggta atcatagaag
     4321 aatcagcaaa gaatccagcc aagggactat cgaacgtcgg gttcaggtat ttgagatgag
     4381 ccctctgtcg ctcaagaacg ctcatgtctg gaaagttcag gagaaggtcg ccaccgatgg
     4441 cgccattcat gctgccgcca ccactgccgc ccgtacgagg gccgggattc tcctccacgt
     4501 caccgcatgt tagaagactt cctctgccct caccgccacc agatccacct ccgctagctc
     4561 ccgcgcctcc tcctcctgag cctcctcctg aggcggcgcc gatcataatt tgggcctccc
     4621 gtgtcctaga gatagccttt gccagcagct cccttgcggt gtcaatatcg acgataggct
     4681 ttgcgtaatt ggtacccaac tctacgccgc ttgccttcaa aacagtcaac ggtgcgtccc
     4741 agggatgatg aatccactcg gtaggcagcc ttgccagctc ggggagccac tggcggatgt
     4801 attccccttc tgggtcgtat ttagccccct gcaaagcggg gttgtcgagc ctatcgagtt
     4861 cgtgcccatc aggaatagat ccggaaatgt actgccaccc aagaatgtcg cactcaaggt
     4921 cagcgtccag cagcgtgtcc cagaagtact tcatgcccca tttccatggg agcaacaaga
     4981 acttgacggc gaaagaagac acaatcactc tgatcctatt atgcatccac cccgtggccc
     5041 acagctccct catgccagcg tcgacaagtg gatacccggt gcgcccctgt cgccaagcct
     5101 tgaatttatc aacatctgcg tcccatggga agaagcgcag gtgagacaag aggctctgtt
     5161 cgtgggtgaa agggaagtta aaacaaatgt agcggctgta ctcccggagt ccgatgcctc
     5221 tcaggaacaa atcggcagac tcctctccct cactgttttt gtctcttgcc cagattatct
     5281 gcttcatccg ggcgcactgg aacacgtgcc gaacagagat ctcgccaaag tgcaggtaag
     5341 ggctcaagag gcttgttgag tttccaacga cttttttgga gttcttagcg tagtcaatca
     5401 gctgcttttc aatgaactcg ttcagaagtt tgtctgcgtt tgaccatcct ggggaccatg
     5461 cgcgagtcag cagggcgtta cttggcttct cagcctcatt ctccagaccc aactcttcga
     5521 tgctacatgc ccagattgcc tcagcggcgg cagtaattgg catcagccgc cagggggggg
     5581 gaagcatcac gctttcgata ctcatgtcca gacatttttt ccagtaagaa ttgaagctag
     5641 tgaaaggctt gcctttctca caataaattt cccatggttc atacaacagg tcgccattgt
     5701 aagactgcac actgatgccc cgctccacca atttctcctt gacggtatgg tcccgcacaa
     5761 ggctaactgg gtcgtagagg tggttgaaaa ccacttttgt tgcgccggtc acccggatgc
     5821 agtccagaat tgcgctaatc gtgttgtggg tcttaatcaa cgtcagatcg gagcccagtg
     5881 ccttgagact ctgtgacagg tgagccaagg actgcttcat ccaccatctg ctggcccggc
     5941 cagggtagaa ctgtccctcc tcctctggac accaaatgaa gacagggaac acagaaccct
     6001 catgggctgc tgctgccaat gcggggttat cttcgattct caggtccctg cgaaaccaga
     6061 caattgtctt cttgtccatc ttcatgctgc cgccaccact gccgccaccg gttgtagcat
     6121 ggggctccgt ggtgatctgt ggctttttac acaagtcctg ataaaattct gattccaaga
     6181 aacgaggata agagttgttc tccatcaggc tgtacaccct cttctgcgct gtggtgaagc
     6241 agccacttgt agcctcttgg atattttggg caatcagagt ttttgtttga aagtctatgt
     6301 ttatctcttt gggagcttcc ttctcgatga agtcggtgta tattttcctt gcttttgagg
     6361 acagtttctg tggtgatttg gttttcttga agtcttcgca agccaaccag aattcaatat
     6421 tttcttcaca gaactcggac tttaaaaacg ctctgaatgc agcaagccca tatttgctgg
     6481 ccagcagttc atcaaatgct tctgcccaga gcagcgcttc ctcaggagaa ggcttgataa
     6541 aagtttgctg tttgcttttc ttgccagttt tgggcttccc aggagtcatg gatcccaagc
     6601 ttgaattcga ctgcgctctc aggcacgaca cgactcctcc gctgcccacc gcagactgag
     6661 gcagcgctga gtcgccggcg ccgcagcgca gatggtcgcg cccgtgcccc cctatctcgc
     6721 gcctcgcgtg gtgcggtccg gctgggccgg cggcggcgcg gacgcgacca aggtggccgg
     6781 gaaggggagt ttgcggggga ccggcgagtg acgtcagcgc gccttcagtg ctgaggcggc
     6841 ggtggcgcgc gccgccaggc gggggcgaag gcactgtccg cggtgctgaa gctggcagtg
     6901 cgcacgcgcc tcgccgcatc ctgtttcccc tccccctctc tgatagggga tgcgcaattt
     6961 ggggaatggg ggttgggtgc ttgtccagtg ggtcggggtc ggtcgtcagg taggcacccc
     7021 caccccgcct catcctggtc ctaaaaccca cttgcactca tacgcagggc cctctgcaga
     7081 gatcttccta gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa
     7141 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     7201 cgggcgacca aaggtcgccc gacgcccggg ctttgcccng ggcggcctca gtgagcgagc
     7261 gagcgcgcag ctggcgt
//
