LOCUS       sequence-484556-        6580 bp    DNA     circular SYN 25-AUG-2025
DEFINITION  The transfer plasmid for packaging AAV expressing chicken OPN5 was 
            constructed by inserting the OPN5 coding sequence into the multiple 
            cloning site under the control of GfaABC1D promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6580)
  AUTHORS   Dai R, Yu T, Weng D, Li H, Cui Y, Wu Z, Guo Q, Zou H, Wu W, Gao X, 
            Qi Z, Ren Y, Wang S, Li Y, Luo M
  TITLE     A neuropsin-based optogenetic tool for precise control of Gq 
            signaling.
  JOURNAL   Sci China Life Sci. 2022 Jul;65(7):1271-1284. doi: 
            10.1007/s11427-022-2122-0. Epub 2022 May 12.
  PUBMED    35579776
REFERENCE   2  (bases 1 to 6580)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 25, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1007/s11427-022-2122-0"; journalName: "Sci China Life Sci"; 
            date: "2022-07"; volume: "65"; issue: "7"; pages: "1271-1284"
COMMENT     Sequence Label: pAAV-GfaABC1D-cOpn5-T2A-EGFP
FEATURES             Location/Qualifiers
     source          1..6580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(137..156)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   454..594
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   454..583
                     /label=AAV2 ITR
     rep_origin      669..1124
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(756..775)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     966..987
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1141..1160)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1260..1282
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1320..1338)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1406..1510
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1511..1579,1580..2371)
                     /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      2542..3130
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3031..3050
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   3192..3321
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             5103..5156
                     /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(5157..5159,5160..5873)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(5157..5159,5160..5870)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5199..5220)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5460..5479)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5807..5828
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5898..6486
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5951..5971)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6369..6380)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    join(6518..6580,1..414)
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
ORIGIN
        1 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc
       61 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc
      121 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc
      181 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg
      241 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt
      301 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg
      361 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt
      421 ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
      481 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
      541 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
      601 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
      661 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
      721 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
      781 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
      841 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
      901 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
      961 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     1021 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     1081 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     1141 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     1201 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     1261 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     1321 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     1381 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     1441 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     1501 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     1561 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     1621 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     1681 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     1741 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     1801 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     1861 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     1921 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     1981 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     2041 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     2101 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     2161 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     2221 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     2281 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     2341 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     2401 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     2461 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     2521 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2581 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2641 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     2701 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2761 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2821 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     2881 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     2941 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3001 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3061 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3121 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3181 tgctcacatg tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc
     3241 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
     3301 actccatcac taggggttcc tgcggccgca cgcgtgatct aacatatcct ggtgtggagt
     3361 aggggacgct gctctgacag aggctcgggg gcctgagctg gctctgtgag ctggggagga
     3421 ggcagacagc caggccttgt ctgcaagcag acctggcagc attgggctgg ccgcccccca
     3481 gggcctcctc ttcatgccca gtgaatgact caccttggca cagacacaat gttcggggtg
     3541 ggcacagtgc ctgcttcccg ccgcacccca gcccccctca aatgccttcc gagaagccca
     3601 ttgagcaggg ggcttgcatt gcaccccagc ctgacagcct ggcatcttgg gataaaagca
     3661 gcacagcccc ctaggggctg cccttgctgt gtggcgccac cggcggtgga gaacaaggct
     3721 ctattcagcc tgtgcccagg aaaggggatc aggggatgcc caggcatgga cagtgggtgg
     3781 caggggggga gaggagggct gtctgcttcc cagaagtcca aggacacaaa tgggtgaggg
     3841 gagagctctc cccatagctg ggctgcggcc caaccccacc ccctcaggct atgccagggg
     3901 gtgttgccag gggcacccgg gcatcgccag tctagcccac tccttcataa agccctcgca
     3961 tcccaggagc gagcagagcc agagcaggtt ggagaggaga cgcatcacct ccgctgctcg
     4021 cggggtctag aatgagtggg atggcatcgg actgcaactc ctcctctcag gaagaatacc
     4081 tcccacacta cgtgcagcag gaagacccct ttgcatcaaa gctgtctaga gaagctgaca
     4141 tcatagcggg gttttatttg acagtaattg ggattctttc aactctggga aacgggtatg
     4201 ttatctttat gtcctcaaag cggaaaaaga agttaagacc cgctgagata atgactgtta
     4261 atttagcggt gtgtgatctg ggcatctcag ttgtaggaaa acccttcagt atcatttcct
     4321 tcttttctca ccgctggatc tttggatgga tgggctgccg ctggtacgga tgggctggct
     4381 tcttctttgg ctgtgggagc cttattacca tgacagccgt cagcctggat agatacttaa
     4441 aaatctgtca cttggcttat ggtacctggc ttaagagaca ccatgccttt atctgcctgg
     4501 cactaatctg ggcctatgcg acgttctggg ctacagtgcc ttttgctggt gtggggagct
     4561 acgctcctga gccgtttggg acctcgtgca ccctggactg gtggctggcg caggcttcag
     4621 tggctgggca ggcttttgtt ctcagcattc ttttcttttg cctcctgttt ccaactgcag
     4681 tgattgtgtt ctcctatgtt aaaattattt taaaagtcaa gtcgtccacc aaagaagttg
     4741 ctcactatga caccaggatc cagaacagcc acatacttga aatgaagctg accaaggtgg
     4801 caatgctgat ctgtgccgga ttcctcattg cttggatccc ctacgccgtg gtctctgtgt
     4861 ggtcagcctt tggacagccg gattccgttc ccattcaatt ttcggtggta ccaactctgc
     4921 ttgccaagtc ggctgccatg tacaacccaa ttatttacca ggtcattgat tgcaaatttg
     4981 cctgctgccg gtcaggtggg ccgaagacac tgcagaagaa gagctctttg aaggaatcga
     5041 ggatgtacac cttatcttca cacagagatt cagcggcact gagcggaacc caactggaag
     5101 tcgagggcag aggaagtctg ctaacatgcg gtgacgtcga ggagaatcct ggcccagtga
     5161 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     5221 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     5281 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     5341 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     5401 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     5461 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     5521 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     5581 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     5641 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     5701 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     5761 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     5821 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taagaattcg
     5881 atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     5941 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     6001 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     6061 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     6121 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     6181 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     6241 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     6301 cctttccttg gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct
     6361 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     6421 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     6481 ccccgcatcg ataccgagcg ctgctcgaga gatctacggg tggcatccct gtgacccctc
     6541 cccagtgcct ctcctggccc tggaagttgc cactccagtg
//
