LOCUS       sequence-265114-        6360 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Viral expression of NpHR3.3-EYFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6360)
  AUTHORS   Fenno LE, Ramakrishnan C, Kim YS, Evans KE, Lo M, Vesuna S, Inoue M,
            Cheung KYM, Yuen E, Pichamoorthy N, Hong ASO, Deisseroth K
  TITLE     Comprehensive Dual- and Triple-Feature Intersectional Single-Vector 
            Delivery of Diverse Functional Payloads to Cells of Behaving 
            Mammals.
  JOURNAL   Neuron. 2020 Jun 20. pii: S0896-6273(20)30432-3. doi: 
            10.1016/j.neuron.2020.06.003.
  PUBMED    32574559
REFERENCE   2  (bases 1 to 6360)
  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"; journalName: "Neuron. 
            2020 Jun 20. pii: S0896-6273(20)30432-3. doi: 
            10.1016/j.neuron.2020.06.003."
COMMENT     Sequence Label: pAAV-nEF-NpHR3.3-EYFP
FEATURES             Location/Qualifiers
     source          1..6360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        188..399
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             412..680
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             1665..1721
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(1722..1724,1725..2435)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1764..1785)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2025..2044)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2372..2393
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2487..3075
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2540..2560)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2958..2969)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3107..3583
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3306..3325)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3623..3763
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3623..3752
                     /label=AAV2 ITR
     rep_origin      3838..4293
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3925..3944)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4135..4156
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4310..4329)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4429..4451
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4489..4507)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4575..4679
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4680..4748,4749..5540)
                     /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(4898..4917)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5711..6299
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6200..6219
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagga tctgcgatcg ctccggtgcc
      181 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
      241 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
      301 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
      361 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc
      421 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc
      481 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta
      541 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc
      601 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct
      661 gttctgcgcc gttacagatc caagctgtga ccggcgccta cggatccgcc accatgacag
      721 agaccctgcc tcccgtgacc gagagtgccg tggcccttca agccgaggtt acccaaaggg
      781 agttgttcga gttcgtgctg aacgaccctt tgcttgcaag cagtctctat atcaacatcg
      841 cacttgcagg actgagtata ctgctgttcg tttttatgac ccgaggactc gatgatccac
      901 gggcaaaact tattgctgtg tcaaccatcc ttgtgcctgt cgtcagcatt gcctcctaca
      961 ctggattggc gagcggcctg acaatttccg ttcttgaaat gccagcgggc cattttgcag
     1021 aaggcagctc agtgatgctg ggaggagaag aggtagatgg tgtagtcacc atgtggggac
     1081 ggtatctcac ctgggcactt tccacgccca tgattctcct cgctctgggt ctcctggccg
     1141 gaagcaatgc tacaaagctc ttcacagcta tcactttcga tatcgctatg tgcgtgactg
     1201 gccttgccgc ggccctgact acctcctccc acctcatgag atggttcttt tacgctatca
     1261 gttgtgcatg ctttctggtg gtcttgtata tcctgctggt ggagtgggca caggacgcca
     1321 aagccgcggg aaccgctgac atgttcaata ccctgaagct gttgacagta gtgatgtggc
     1381 tggggtatcc aattgtgtgg gctcttggag tcgagggtat cgcggtgttg cccgttgggg
     1441 tgacgagctg gggatattct ttcctggata tcgtggcaaa gtacattttc gcattcttgc
     1501 tcctgaacta tctgacgtca aacgaatctg tcgtgtccgg cagcattttg gatgttccat
     1561 ctgcttctgg gaccccggct gatgataaga gcaggatcac cagcgagggc gagtacatcc
     1621 ccctggacca gatcgacatc aacgtgggcg cgcccggctc cggagccacg aacttctctc
     1681 tgttaaagca agcaggagac gtggaagaaa accccggtcc cgtgagcaag ggcgaggagc
     1741 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     1801 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     1861 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ttcggctacg
     1921 gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     1981 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     2041 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     2101 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     2161 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     2221 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     2281 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccgccc
     2341 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     2401 ccgggatcac tctcggcatg gacgagctgt acaagttctg ctacgagaac gaggtgtaat
     2461 gagaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     2521 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     2581 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     2641 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     2701 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     2761 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     2821 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     2881 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     2941 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     3001 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     3061 gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctacgggt ggcatccctg
     3121 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct
     3181 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat
     3241 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg
     3301 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc
     3361 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc
     3421 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt
     3481 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt
     3541 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt tgtaggtaac
     3601 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     3661 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     3721 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     3781 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     3841 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3901 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3961 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4021 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     4081 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4141 tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag
     4201 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4261 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     4321 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     4381 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     4441 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     4501 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     4561 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4621 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4681 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4741 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4801 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4861 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4921 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4981 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5041 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5101 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5161 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5221 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5281 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5341 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5401 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5461 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5521 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5581 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5641 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5701 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5761 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5821 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     5881 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5941 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6001 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6061 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6121 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6181 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6241 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6301 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
