LOCUS       sequence-196840-        7058 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of eNpHR 3.0 fused to EYFP driven by CaMKIIa promoter
            for optogenetic inhibition.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7058)
  TITLE     eNpHR3.0-EYFP
REFERENCE   2  (bases 1 to 7058)
  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"
COMMENT     Sequence Label: pAAV-CaMKIIa-eNpHR 3.0-EYFP
FEATURES             Location/Qualifiers
     source          1..7058
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   54..183
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        202..1490
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     CDS             join(2473..2475,2476..3186)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2515..2536)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2776..2795)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3123..3144
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3238..3826
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3291..3311)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3709..3720)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3858..4334
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4057..4076)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4374..4514
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4374..4503
                     /label=AAV2 ITR
     rep_origin      4589..5044
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4676..4695)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4886..4907
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5061..5080)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5180..5202
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5240..5258)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5326..5430
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5431..5499,5500..6291)
                     /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(5649..5668)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6462..7050
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6951..6970
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtcctgcag
       61 gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc
      121 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
      181 cctgcggccg cacgcgttta acattatggc cttaggtcac ttcatctcca tggggttctt
      241 cttctgattt tctagaaaat gagatggggg tgcagagagc ttcctcagtg acctgcccag
      301 ggtcacatca gaaatgtcag agctagaact tgaactcaga ttactaatct taaattccat
      361 gccttggggg catgcaagta cgatatacag aaggagtgaa ctcattaggg cagatgacca
      421 atgagtttag gaaagaagag tccagggcag ggtacatcta caccacccgc ccagccctgg
      481 gtgagtccag ccacgttcac ctcattatag ttgcctctct ccagtcctac cttgacggga
      541 agcacaagca gaaactggga caggagcccc aggagaccaa atcttcatgg tccctctggg
      601 aggatgggtg gggagagctg tggcagaggc ctcaggaggg gccctgctgc tcagtggtga
      661 cagatagggg tgagaaagca gacagagtca ttccgtcagc attctgggtc tgtttggtac
      721 ttcttctcac gctaaggtgg cggtgtgata tgcacaatgg ctaaaaagca gggagagctg
      781 gaaagaaaca aggacagaga cagaggccaa gtcaaccaga ccaattccca gaggaagcaa
      841 agaaaccatt acagagacta caagggggaa gggaaggaga gatgaattag cttcccctgt
      901 aaaccttaga acccagctgt tgccagggca acggggcaat acctgtctct tcagaggaga
      961 tgaagttgcc agggtaacta catcctgtct ttctcaagga ccatcccaga atgtggcacc
     1021 cactagccgt taccatagca actgcctctt tgccccactt aatcccatcc cgtctgttaa
     1081 aagggcccta tagttggagg tgggggaggt aggaagagcg atgatcactt gtggactaag
     1141 tttgttcgca tccccttctc caaccccctc agtacatcac cctgggggaa cagggtccac
     1201 ttgctcctgg gcccacacag tcctgcagta ttgtgtatat aaggccaggg caaagaggag
     1261 caggttttaa agtgaaaggc aggcaggtgt tggggaggca gttaccgggg caacgggaac
     1321 agggcgtttc ggaggtggtt gccatgggga cctggatgct gacgaaggct cgcgaggctg
     1381 tgagcagcca cagtgccctg ctcagaagcc ccaagctcgt cagtcaagcc ggttctccgt
     1441 ttgcactcag gagcacgggc aggcgagtgg cccctagttc tgggggcagc tctagagcgg
     1501 taccggatcc ccgggtaccg gtaggccacc atgacagaga ccctgcctcc cgtgaccgag
     1561 agtgccgtgg cccttcaagc cgaggttacc caaagggagt tgttcgagtt cgtgctgaac
     1621 gaccctttgc ttgcaagcag tctctatatc aacatcgcac ttgcaggact gagtatactg
     1681 ctgttcgttt ttatgacccg aggactcgat gatccacggg caaaacttat tgctgtgtca
     1741 accatccttg tgcctgtcgt cagcattgcc tcctacactg gattggcgag cggcctgaca
     1801 atttccgttc ttgaaatgcc agcgggccat tttgcagaag gcagctcagt gatgctggga
     1861 ggagaagagg tagatggtgt agtcaccatg tggggacggt atctcacctg ggcactttcc
     1921 acgcccatga ttctcctcgc tctgggtctc ctggccggaa gcaatgctac aaagctcttc
     1981 acagctatca ctttcgatat cgctatgtgc gtgactggcc ttgccgcggc cctgactacc
     2041 tcctcccacc tcatgagatg gttctggtac gctatcagtt gtgcatgctt tctggtggtc
     2101 ttgtatatcc tgctggtgga gtgggcacag gacgccaaag ccgcgggaac cgctgacatg
     2161 ttcaataccc tgaagctgtt gacagtagtg atgtggctgg ggtatccaat tgtgtgggct
     2221 cttggagtcg agggtatcgc ggtgttgccc gttggggtga cgagctgggg atattctttc
     2281 ctggatatcg tggcaaagta cattttcgca ttcttgctcc tgaactatct gacgtcaaac
     2341 gaatctgtcg tgtccggcag cattttggat gttccatctg cttctgggac cccggctgat
     2401 gatgcggccg ccaagagcag gatcaccagc gagggcgagt acatccccct ggaccagatc
     2461 gacatcaacg tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     2521 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     2581 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     2641 tggcccaccc tcgtgaccac cttcggctac ggcctgcagt gcttcgcccg ctaccccgac
     2701 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     2761 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     2821 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     2881 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     2941 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     3001 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     3061 gacaaccact acctgagcta ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     3121 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     3181 tacaagttct gctacgagaa cgaggtgtaa tgagaattcg atatcaagct tatcgataat
     3241 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     3301 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     3361 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     3421 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     3481 tggggcattg ccaccacctg tcagctcctt tccggaactt tcgctttccc cctccctatt
     3541 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     3601 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     3661 tatgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     3721 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     3781 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgagcg
     3841 ctgctcgaga gatctacggg tggcatccct gtgacccctc cccagtgcct ctcctggccc
     3901 tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt
     3961 tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg
     4021 ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg
     4081 cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc
     4141 ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt
     4201 ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc
     4261 aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc
     4321 ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc
     4381 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     4441 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     4501 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4561 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     4621 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct
     4681 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4741 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4801 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4861 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4921 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
     4981 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     5041 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     5101 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     5161 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     5221 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     5281 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5341 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5401 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5461 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5521 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5581 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5641 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5701 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5761 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5821 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5881 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5941 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     6001 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6061 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6121 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6181 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6241 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6301 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6361 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6421 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6481 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6541 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6601 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6661 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6721 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6781 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6841 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6901 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6961 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7021 tgctcgtcag gggggcggag cctatggaaa aacgccag
//
