LOCUS       sequence-461235-        8370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV encoding the 173 amino acid version PiGM-Ii with minimal RGS10 
            domain, CRY2 (535 amino acid version), CIB1 (81 amino acid 
            version)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8370)
  TITLE     PiGM-Iq system plasmids
REFERENCE   2  (bases 1 to 8370)
  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-Ii (173, 535/81, eGFP)
FEATURES             Location/Qualifiers
     source          1..8370
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2..21)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     248..268
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(277..705)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     832..854
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     864..883
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        864..882
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(970..988)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(971..988)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    1002..1018
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1026..1032,1033..1050,1051..1056))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1071..1092
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(1381..1969)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1461..1480)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2140..2931,2932..3000))
                     /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     2763..2782
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3001..3105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3131..3586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     repeat_region   3658..3787
                     /label=AAV2 ITR
     promoter        3869..4317
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             6522..6575
                     /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(6867..6869,6870..6872,6873..7583)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6867..6869,6870..6872,6873..7583)
                     /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(6912..6933)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7173..7192)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7520..7541
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7738..7758)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   8185..8314
                     /label=AAV2 ITR
ORIGIN
        1 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
       61 ctgggcggag ttaggggcgg gatgggcgga gktaggggsg ggastatggt tgstgactaa
      121 wtgrgatgca tgctttgcat acttctgcct gctggggrgc ctggggactt tccacacctg
      181 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
      241 tttccacacc ctaactgaca cacattccac agaattaatt cgcgttaaat ttttgttaaa
      301 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
      361 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
      421 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
      481 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
      541 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
      601 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
      661 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca
      721 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
      781 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
      841 gacgttgtaa aacgacggcc aggtaatacg actcactata gggcgagaat tcggtctsav
      901 hrvgcaatam tymwartcaw tggcngcaag cttygwtmag gatdangnhs bhtgagacca
      961 agcttcactt ctatagtgtc acctaaatag ctcgtgtgaa attgttatcc gctcacaatt
     1021 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     1081 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     1141 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tngggcgctc
     1201 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     1261 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     1321 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1381 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     1441 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     1501 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     1561 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     1621 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     1681 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     1741 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     1801 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     1861 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     1921 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     1981 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2041 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2101 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2161 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     2221 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     2281 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     2341 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     2401 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     2461 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     2521 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     2581 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     2641 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     2701 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     2761 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     2821 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     2881 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     2941 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     3001 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     3061 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     3121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     3181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     3241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     3301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     3361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     3421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     3481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     3541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
     3601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     3661 cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt
     3721 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg
     3781 ggttccttgt agttaatgat taacccgcca tgctacttat ctacgtagcc atgctctagg
     3841 aagatctctg cagagggccc tgcgtatgag tgcaagtggg ttttaggacc aggatgaggc
     3901 ggggntgggg gtgcctacct gacgaccgac cccgacccac tggacaagca cccaaccccc
     3961 attccccaaa ttgcgcatcc cctatcagag agggggaggg gaaacaggat gcggcgaggc
     4021 gcgtgcgcac tgccagcttc agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg
     4081 cgccaccgcc gcctcagcac tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact
     4141 ccccttcccg gccaccttgg tcgcgtccgc gccgccgccg gcccagccgg accgcaccac
     4201 gcgaggcgcg agataggggg gcacgggcgc gaccatctgc gctgcggcgc cggcgactca
     4261 gcgctgcctc agtctgcggt gggcagcgga ggagtcgtgt cgtgcctgag agcgcagtcg
     4321 aattcaagct tatgttcacc cgcgccgtga gccgactgag caggaagcgg ccgccgtctg
     4381 atatccacga cggcgatggg agctcaagca gcggccacca gagcctcaag agcacagcca
     4441 agtgggcgtc ctccctggag aatcttctgg aagatccaga gggcgtgaag agattcaggg
     4501 aatttctgaa aaaggaattc agtgaggaaa acgtcttgtt ttggctagcg tgtgaagatt
     4561 tcaagaaaac ggaggacaag aagcagatgc aggaaaaggc caagaagatc tacatgacct
     4621 tcctgtccaa taaggcctct tcacaagtca atgtggaggg gcagtctcgg ctcactgaaa
     4681 agattctgga agaaccacac cctctgatgt tccaaaagct ccaggaccag atcttcaatc
     4741 tcatgaagta tgacagctac agccgcttct tgaagtctga cttgtttctg aagcaccggc
     4801 ggactgagga agaggaagag gatcctccgg acgctcaaac tgcagctacc ggtggcggca
     4861 gcatgaagat ggacaagaag acaattgtct ggtttcgcag ggacctgaga atcgaagata
     4921 accccgcatt ggcagcagca gcccatgagg gttctgtgtt ccctgtcttc atttggtgtc
     4981 cagaggagga gggacagttc taccctggcc gggccagcag atggtggatg aagcagtcct
     5041 tggctcacct gtcacagagt ctcaaggcac tgggctccga tctgacgttg attaagaccc
     5101 acaacacgat tagcgcaatt ctggactgca tccgggtgac cggcgcaaca aaagtggttt
     5161 tcaaccacct ctacgaccca gttagccttg tgcgggacca taccgtcaag gagaaattgg
     5221 tggagcgggg catcagtgtg cagtcttaca atggcgacct gttgtatgaa ccatgggaaa
     5281 tttattgtga gaaaggcaag cctttcacta gcttcaattc ttactggaaa aaatgtctgg
     5341 acatgagtat cgaaagcgtg atgcttcccc ccccctggcg gctgatgcca attactgccg
     5401 ccgctgaggc aatctgggca tgtagcatcg aagagttggg tctggagaat gaggctgaga
     5461 agccaagtaa cgccctgctg actcgcgcat ggtccccagg atggtcaaac gcagacaaac
     5521 ttctgaacga gttcattgaa aagcagctga ttgactacgc taagaactcc aaaaaagtcg
     5581 ttggaaactc aacaagcctc ttgagccctt acctgcactt tggcgagatc tctgttcggc
     5641 acgtgttcca gtgcgcccgg atgaagcaga taatctgggc aagagacaaa aacagtgagg
     5701 gagaggagtc tgccgatttg ttcctgagag gcatcggact ccgggagtac agccgctaca
     5761 tttgttttaa cttccctttc acccacgaac agagcctctt gtctcacctg cgcttcttcc
     5821 catgggacgc agatgttgat aaattcaagg cttggcgaca ggggcgcacc gggtatccac
     5881 ttgtcgacgc tggcatgagg gagctgtggg ccacggggtg gatgcataat aggatcagag
     5941 tgattgtgtc ttctttcgcc gtcaagttct tgttgctccc atggaaatgg ggcatgaagt
     6001 acttctggga cacgctgctg gacgctgacc ttgagtgcga cattcttggg tggcagtaca
     6061 tttccggatc tattcctgat gggcacgaac tcgataggct cgacaacccc gctttgcagg
     6121 gggctaaata cgacccagaa ggggaataca tccgccagtg gctccccgag ctggcaaggc
     6181 tgcctaccga gtggattcat catccctggg acgcaccgtt gactgttttg aaggcaagcg
     6241 gcgtagagtt gggtaccaat tacgcaaagc ctatcgtcga tattgacacc gcaagggagc
     6301 tgctggcaaa ggctatctct aggacacggg aggcccaaat tatgatcggc gccgcccctg
     6361 atgagattgt agcagatagc ttcgaggcct taggggctaa taccattaaa gaacctggtc
     6421 tttgcccatc tgtgtcttct aatgaccaac aagtaccttc ggctgtcgat cttaagttgg
     6481 ctggacttga tattgctagc ggaggtggat ctggtggcgg tgagggcaga ggaagtcttc
     6541 taacatgcgg tgacgtggag gagaatcccg gccctcgtac gggcggcagt ggtggcggca
     6601 gcatgaatgg cgccatcggt ggcgaccttc tcctgaactt tccagacatg agcgttcttg
     6661 agcgacagag ggctcatctc aaatacctga acccgacgtt cgatagtccc ttggctggat
     6721 tctttgctga ttcttctatg attaccggcg gcgaaatgga ttcttatctg agtactgctg
     6781 gtttgaacct gcctatgatg tacggagaga ccacagtgga gggagatagc agactgtcca
     6841 tctccggagg aggcgcggga atcgatatgg tgagcaaggg cgaggagctg ttcaccgggg
     6901 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     6961 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     7021 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     7081 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     7141 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     7201 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     7261 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     7321 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     7381 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     7441 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     7501 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     7561 tcggcatgga cgagctgtat aagggtaaaa agaagaaaaa gaagtcaaag acaaagtgtg
     7621 taattatgta agaattctgc agatatccat cacactggcg gccgctcgag catgcatcta
     7681 gaataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     7741 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     7801 ccgtatggct ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga
     7861 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     7921 ttccgtggtg tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccatcactag
     7981 ttaggcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     8041 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg
     8101 tgggaggttt tttaaaatcg ataaggatct tcctagagca tggctacgta gataagtagc
     8161 atggcgggtt aatcattaac tacaaggaac ccctagtgat ggagttggcc actccctctc
     8221 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     8281 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcat taatgaatcg gccaacgcgc
     8341 ggggagaggc ggtttgcgta ttgggcgctc
//
