LOCUS       sequence-481804-        6574 bp    DNA     circular SYN 04-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 6574)
  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 6574)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 4, 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-mcherry
FEATURES             Location/Qualifiers
     source          1..6574
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..590
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     491..510
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   652..781
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             2563..2616
                     /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             2617..3327
                     /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(2762..2780)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3030..3049
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3352..3940
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3405..3425)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3823..3834)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3972..4448
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4171..4190)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4488..4628
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4488..4617
                     /label=AAV2 ITR
     rep_origin      4703..5158
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4790..4809)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5000..5021
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5175..5194)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5294..5316
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5354..5372)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5440..5544
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5545..5613,5614..6405)
                     /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"
ORIGIN
        1 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
       61 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
      121 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
      181 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
      241 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
      301 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
      361 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
      421 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
      481 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
      541 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
      601 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc
      661 agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt tggtcgcccg
      721 gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc
      781 tgcggccgca cgcgtgatct aacatatcct ggtgtggagt aggggacgct gctctgacag
      841 aggctcgggg gcctgagctg gctctgtgag ctggggagga ggcagacagc caggccttgt
      901 ctgcaagcag acctggcagc attgggctgg ccgcccccca gggcctcctc ttcatgccca
      961 gtgaatgact caccttggca cagacacaat gttcggggtg ggcacagtgc ctgcttcccg
     1021 ccgcacccca gcccccctca aatgccttcc gagaagccca ttgagcaggg ggcttgcatt
     1081 gcaccccagc ctgacagcct ggcatcttgg gataaaagca gcacagcccc ctaggggctg
     1141 cccttgctgt gtggcgccac cggcggtgga gaacaaggct ctattcagcc tgtgcccagg
     1201 aaaggggatc aggggatgcc caggcatgga cagtgggtgg caggggggga gaggagggct
     1261 gtctgcttcc cagaagtcca aggacacaaa tgggtgaggg gagagctctc cccatagctg
     1321 ggctgcggcc caaccccacc ccctcaggct atgccagggg gtgttgccag gggcacccgg
     1381 gcatcgccag tctagcccac tccttcataa agccctcgca tcccaggagc gagcagagcc
     1441 agagcaggtt ggagaggaga cgcatcacct ccgctgctcg cggggtctag aatgagtggg
     1501 atggcatcgg actgcaactc ctcctctcag gaagaatacc tcccacacta cgtgcagcag
     1561 gaagacccct ttgcatcaaa gctgtctaga gaagctgaca tcatagcggg gttttatttg
     1621 acagtaattg ggattctttc aactctggga aacgggtatg ttatctttat gtcctcaaag
     1681 cggaaaaaga agttaagacc cgctgagata atgactgtta atttagcggt gtgtgatctg
     1741 ggcatctcag ttgtaggaaa acccttcagt atcatttcct tcttttctca ccgctggatc
     1801 tttggatgga tgggctgccg ctggtacgga tgggctggct tcttctttgg ctgtgggagc
     1861 cttattacca tgacagccgt cagcctggat agatacttaa aaatctgtca cttggcttat
     1921 ggtacctggc ttaagagaca ccatgccttt atctgcctgg cactaatctg ggcctatgcg
     1981 acgttctggg ctacagtgcc ttttgctggt gtggggagct acgctcctga gccgtttggg
     2041 acctcgtgca ccctggactg gtggctggcg caggcttcag tggctgggca ggcttttgtt
     2101 ctcagcattc ttttcttttg cctcctgttt ccaactgcag tgattgtgtt ctcctatgtt
     2161 aaaattattt taaaagtcaa gtcgtccacc aaagaagttg ctcactatga caccaggatc
     2221 cagaacagcc acatacttga aatgaagctg accaaggtgg caatgctgat ctgtgccgga
     2281 ttcctcattg cttggatccc ctacgccgtg gtctctgtgt ggtcagcctt tggacagccg
     2341 gattccgttc ccattcaatt ttcggtggta ccaactctgc ttgccaagtc ggctgccatg
     2401 tacaacccaa ttatttacca ggtcattgat tgcaaatttg cctgctgccg gtcaggtggg
     2461 ccgaagacac tgcagaagaa gagctctttg aaggaatcga ggatgtacac cttatcttca
     2521 cacagagatt cagcggcact gagcggaacc caactggaag tcgagggcag aggaagtctg
     2581 ctaacatgcg gtgacgtcga ggagaatcct ggcccaatgg tgagcaaggg cgaggaggat
     2641 aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac
     2701 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     2761 gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct
     2821 cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg
     2881 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
     2941 gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag
     3001 ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg
     3061 gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag
     3121 aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc
     3181 aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc
     3241 cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc
     3301 ggcggcatgg acgagctgta caagtaagaa ttcgatatca agcttatcga taatcaacct
     3361 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     3421 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     3481 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     3541 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     3601 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     3661 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     3721 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt
     3781 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     3841 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     3901 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg agcgctgctc
     3961 gagagatcta cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag
     4021 ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg
     4081 actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag
     4141 ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg
     4201 cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc
     4261 ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt
     4321 ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga
     4381 tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc
     4441 ctgtccttct gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt
     4501 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     4561 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     4621 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     4681 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4741 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct
     4801 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4861 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     4921 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4981 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
     5041 tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat
     5101 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     5161 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     5221 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     5281 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     5341 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     5401 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5461 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5521 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5581 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5641 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5701 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5761 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5821 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5881 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5941 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     6001 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     6061 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     6121 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     6181 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6241 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6301 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6361 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6421 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6481 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6541 gcgtcagacc ccgtagaaaa gatcaaagga tctt
//
