LOCUS       sequence-272743-        6349 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 6349)
  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 6349)
  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..6349
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        177..388
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             401..669
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             1654..1710
                     /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(1711..1713,1714..2424)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1753..1774)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2014..2033)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2361..2382
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2476..3064
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2529..2549)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2947..2958)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3096..3572
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3295..3314)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3612..3752
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3612..3741
                     /label=AAV2 ITR
     rep_origin      3827..4282
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3914..3933)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4124..4145
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4299..4318)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4418..4440
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4478..4496)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4564..4668
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4669..4737,4738..5529)
                     /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(4887..4906)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5700..6288
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6189..6208
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaaggat ctgcgatcgc tccggtgccc gtcagtgggc
      181 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg
      241 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
      301 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
      361 tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc
      421 acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc
      481 tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc
      541 gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac
      601 gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg
      661 ttacagatcc aagctgtgac cggcgcctac ggatccgcca ccatgacaga gaccctgcct
      721 cccgtgaccg agagtgccgt ggcccttcaa gccgaggtta cccaaaggga gttgttcgag
      781 ttcgtgctga acgacccttt gcttgcaagc agtctctata tcaacatcgc acttgcagga
      841 ctgagtatac tgctgttcgt ttttatgacc cgaggactcg atgatccacg ggcaaaactt
      901 attgctgtgt caaccatcct tgtgcctgtc gtcagcattg cctcctacac tggattggcg
      961 agcggcctga caatttccgt tcttgaaatg ccagcgggcc attttgcaga aggcagctca
     1021 gtgatgctgg gaggagaaga ggtagatggt gtagtcacca tgtggggacg gtatctcacc
     1081 tgggcacttt ccacgcccat gattctcctc gctctgggtc tcctggccgg aagcaatgct
     1141 acaaagctct tcacagctat cactttcgat atcgctatgt gcgtgactgg ccttgccgcg
     1201 gccctgacta cctcctccca cctcatgaga tggttctttt acgctatcag ttgtgcatgc
     1261 tttctggtgg tcttgtatat cctgctggtg gagtgggcac aggacgccaa agccgcggga
     1321 accgctgaca tgttcaatac cctgaagctg ttgacagtag tgatgtggct ggggtatcca
     1381 attgtgtggg ctcttggagt cgagggtatc gcggtgttgc ccgttggggt gacgagctgg
     1441 ggatattctt tcctggatat cgtggcaaag tacattttcg cattcttgct cctgaactat
     1501 ctgacgtcaa acgaatctgt cgtgtccggc agcattttgg atgttccatc tgcttctggg
     1561 accccggctg atgataagag caggatcacc agcgagggcg agtacatccc cctggaccag
     1621 atcgacatca acgtgggcgc gcccggctcc ggagccacga acttctctct gttaaagcaa
     1681 gcaggagacg tggaagaaaa ccccggtccc gtgagcaagg gcgaggagct gttcaccggg
     1741 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     1801 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     1861 ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct tcggctacgg cctgcagtgc
     1921 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     1981 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     2041 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     2101 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     2161 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     2221 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     2281 ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccgccct gagcaaagac
     2341 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     2401 ctcggcatgg acgagctgta caagttctgc tacgagaacg aggtgtaatg agaattcgat
     2461 atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     2521 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     2581 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     2641 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     2701 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     2761 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     2821 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     2881 tttccttggc tgctcgccta tgttgccacc tggattctgc gcgggacgtc cttctgctac
     2941 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     3001 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     3061 ccgcatcgat accgagcgct gctcgagaga tctacgggtg gcatccctgt gacccctccc
     3121 cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa
     3181 aattaagttg catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg
     3241 gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt
     3301 gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg
     3361 ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca
     3421 ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg
     3481 tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac
     3541 aggcgtgaac cactgctccc ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac
     3601 cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     3661 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     3721 agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac
     3781 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     3841 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3901 gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3961 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4021 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     4081 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4141 actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg
     4201 atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     4261 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
     4321 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     4381 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     4441 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     4501 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     4561 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4621 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4681 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4741 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     4801 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     4861 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     4921 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     4981 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5041 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     5101 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5161 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     5221 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     5281 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5341 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5401 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     5461 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     5521 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     5581 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5641 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5701 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5761 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5821 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     5881 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5941 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6001 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6061 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6121 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6181 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6241 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6301 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
