LOCUS       sequence-484554-        7161 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 a EF1-alpha promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7161)
  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 7161)
  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-EF1-alpha-cOpn5-T2A-mcherry
FEATURES             Location/Qualifiers
     source          1..7161
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     347..366
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   508..637
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        738..1916
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          969..1907
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1864..1884
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3006..3059
                     /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             3060..3770
                     /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(3205..3223)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3473..3492
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3795..4383
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3848..3868)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4266..4277)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4415..4891
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4614..4633)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4931..5071
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4931..5060
                     /label=AAV2 ITR
     rep_origin      5146..5601
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5233..5252)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5443..5464
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5618..5637)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5737..5759
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5797..5815)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5883..5987
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5988..6056,6057..6848)
                     /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(6206..6225)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7019..7161,1..446)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
       61 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
      121 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
      181 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
      241 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
      301 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
      361 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
      421 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
      481 ccttttgctg gccttttgct cacatgtcct gcaggcagct gcgcgctcgc tcgctcactg
      541 aggccgcccg ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc
      601 agagagggag tggccaactc catcactagg ggttcctgcg gccgcacgcg taagctttgc
      661 aaagatggat aaagttttaa acagagagga atctttgcag ctaatggacc ttctaggtct
      721 tgaaaggagt gggaattggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
      781 tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg
      841 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
      901 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
      961 gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc
     1021 cttgcgtgcc ttgaattact tccacctggc tgcagtacgt gattcttgat cccgagcttc
     1081 gggttggaag tgggtgggag agttcgaggc cttgcgctta aggagcccct tcgcctcgtg
     1141 cttgagttga ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc
     1201 gcgcctgtct cgctgctttc gataagtctc tagccattta aaatttttga tgacctgctg
     1261 cgacgctttt tttctggcaa gatagtcttg taaatgcggg ccaagatctg cacactggta
     1321 tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg
     1381 cgaggcgggg cctgcgagcg cggccaccga gaatcggacg ggggtagtct caagctggcc
     1441 ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc
     1501 tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag
     1561 ggagctcaaa atggaggacg cggcgctcgg gagagcgggc gggtgagtca cccacacaaa
     1621 ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga ctccacggag taccgggcgc
     1681 cgtccaggca cctcgattag ttctcgagct tttggagtac gtcgtcttta ggttgggggg
     1741 aggggtttta tgcgatggag tttccccaca ctgagtgggt ggagactgaa gttaggccag
     1801 cttggcactt gatgtaattc tccttggaat ttgccctttt tgagtttgga tcttggttca
     1861 ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc atttcaggtg tcgtgaggta
     1921 ccggatcctc tagaatgagt gggatggcat cggactgcaa ctcctcctct caggaagaat
     1981 acctcccaca ctacgtgcag caggaagacc cctttgcatc aaagctgtct agagaagctg
     2041 acatcatagc ggggttttat ttgacagtaa ttgggattct ttcaactctg ggaaacgggt
     2101 atgttatctt tatgtcctca aagcggaaaa agaagttaag acccgctgag ataatgactg
     2161 ttaatttagc ggtgtgtgat ctgggcatct cagttgtagg aaaacccttc agtatcattt
     2221 ccttcttttc tcaccgctgg atctttggat ggatgggctg ccgctggtac ggatgggctg
     2281 gcttcttctt tggctgtggg agccttatta ccatgacagc cgtcagcctg gatagatact
     2341 taaaaatctg tcacttggct tatggtacct ggcttaagag acaccatgcc tttatctgcc
     2401 tggcactaat ctgggcctat gcgacgttct gggctacagt gccttttgct ggtgtgggga
     2461 gctacgctcc tgagccgttt gggacctcgt gcaccctgga ctggtggctg gcgcaggctt
     2521 cagtggctgg gcaggctttt gttctcagca ttcttttctt ttgcctcctg tttccaactg
     2581 cagtgattgt gttctcctat gttaaaatta ttttaaaagt caagtcgtcc accaaagaag
     2641 ttgctcacta tgacaccagg atccagaaca gccacatact tgaaatgaag ctgaccaagg
     2701 tggcaatgct gatctgtgcc ggattcctca ttgcttggat cccctacgcc gtggtctctg
     2761 tgtggtcagc ctttggacag ccggattccg ttcccattca attttcggtg gtaccaactc
     2821 tgcttgccaa gtcggctgcc atgtacaacc caattattta ccaggtcatt gattgcaaat
     2881 ttgcctgctg ccggtcaggt gggccgaaga cactgcagaa gaagagctct ttgaaggaat
     2941 cgaggatgta caccttatct tcacacagag attcagcggc actgagcgga acccaactgg
     3001 aagtcgaggg cagaggaagt ctgctaacat gcggtgacgt cgaggagaat cctggcccaa
     3061 tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg
     3121 tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
     3181 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct
     3241 tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc
     3301 ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg
     3361 tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg
     3421 gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa
     3481 tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg
     3541 ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg
     3601 aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca
     3661 acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac
     3721 gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtaa gaattcgata
     3781 tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     3841 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     3901 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     3961 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     4021 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     4081 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     4141 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     4201 ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     4261 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     4321 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     4381 cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc
     4441 agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa
     4501 attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg
     4561 ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg
     4621 ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt
     4681 tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag
     4741 gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt
     4801 ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca
     4861 ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc
     4921 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     4981 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     5041 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     5101 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     5161 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     5221 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     5281 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     5341 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     5401 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     5461 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     5521 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     5581 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     5641 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     5701 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     5761 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     5821 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     5881 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     5941 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     6001 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     6061 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     6121 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     6181 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     6241 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     6301 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     6361 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     6421 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     6481 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     6541 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     6601 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     6661 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     6721 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     6781 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     6841 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     6901 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     6961 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     7021 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     7081 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     7141 gcagagcgca gataccaaat a
//
