LOCUS       sequence-194452-        6989 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of ChR2 fused to YFP driven byEF1a promoter for 
            optogenetic activation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6989)
  TITLE     Deisseroth AAV plasmids
REFERENCE   2  (bases 1 to 6989)
  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.EF1a.ChR2-YFP.WPRE.hGH
FEATURES             Location/Qualifiers
     source          1..6989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   38..167
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        236..1414
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          467..1405
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1362..1382
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1430..1439
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1436..2362
                     /codon_start=1
                     /product="N-terminal fragment of channelrhodopsin-2 from 
                     the alga Chlamydomonas reinhardtii (Nagel et al., 2003)"
                     /label=ChR2
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIHLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     regulatory      2372..2381
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2378..2380,2381..2383,2384..3097)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2423..2444)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2684..2703)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3031..3052
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3120..3708
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3173..3193)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3591..3602)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3773..4249
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3972..3991)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4289..4429
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4289..4418
                     /label=AAV2 ITR
     rep_origin      4504..4959
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4591..4610)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4801..4822
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4976..4995)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5095..5117
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5155..5173)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5241..5345
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5346..5414,5415..6206)
                     /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(5564..5583)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6377..6965
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6866..6885
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggttcctgg ccttttgctg gccttttgct cacatgtcct gcaggcagct gcgcgctcgc
       61 tcgctcactg aggccgcccg ggcgtcgggc gacctttggt cgcccggcct cagtgagcga
      121 gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctgcg gccgcacgcg
      181 tgagaggaat ctttgcagct aatggacctt ctaggtcttg aaaggagtgg gaattggctc
      241 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
      301 ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
      361 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
      421 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg
      481 tggttcccgc gggcctggcc tctttacggg ttatggccct tgcgtgcctt gaattacttc
      541 cacctggctg cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag
      601 ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct tgagttgagg cctggcctgg
      661 gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga
      721 taagtctcta gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga
      781 tagtcttgta aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg
      841 cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg
      901 gccaccgaga atcggacggg ggtagtctca agctggccgg cctgctctgg tgcctggcct
      961 cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg caccagttgc
     1021 gtgagcggaa agatggccgc ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg
     1081 gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc
     1141 agccgtcgct tcatgtgact ccacggagta ccgggcgccg tccaggcacc tcgattagtt
     1201 ctcgagcttt tggagtacgt cgtctttagg ttggggggag gggttttatg cgatggagtt
     1261 tccccacact gagtgggtgg agactgaagt taggccagct tggcacttga tgtaattctc
     1321 cttggaattt gccctttttg agtttggatc ttggttcatt ctcaagcctc agacagtggt
     1381 tcaaagtttt tttcttccat ttcaggtgtc gtgaggaatt ctctaggctg ccaccatgga
     1441 ttatggaggc gccctgagtg ccgttgggcg cgagctgcta tttgtaacga acccagtagt
     1501 cgtcaatggc tctgtacttg tgcctgagga ccagtgttac tgcgcgggct ggattgagtc
     1561 gcgtggcaca aacggtgccc aaacggcgtc gaacgtgctg caatggcttg ctgctggctt
     1621 ctccatccta ctgcttatgt tttacgccta ccaaacatgg aagtcaacct gcggctggga
     1681 ggagatctat gtgtgcgcta tcgagatggt caaggtgatt ctcgagttct tcttcgagtt
     1741 taagaacccg tccatgctgt atctagccac aggccaccgc gtccagtggt tgcgttacgc
     1801 cgagtggctt ctcacctgcc cggtcattct cattcacctg tcaaacctga cgggcttgtc
     1861 caacgactac agcaggcgca ccatgggtct gcttgtgtct gatattggca caattgtgtg
     1921 gggcgccact tccgccatgg ccaccggata cgtcaaggtc atcttcttct gcctgggtct
     1981 gtgttatggt gctaacacgt tctttcacgc tgccaaggcc tacatcgagg gttaccacac
     2041 cgtgccgaag ggccggtgtc gccaggtggt gactggcatg gcttggctct tcttcgtatc
     2101 atggggtatg ttccccatcc tgttcatcct cggccccgag ggcttcggcg tcctgagcgt
     2161 gtacggctcc accgtcggcc acaccatcat tgacctgatg tcgaagaact gctggggtct
     2221 gctcggccac tacctgcgcg tgctgatcca cgagcatatc ctcatccacg gcgacattcg
     2281 caagaccacc aaattgaaca ttggtggcac tgagattgag gtcgagacgc tggtggagga
     2341 cgaggccgag gctggcgcgg tacccgcggc cgccaccatg gtgagcaagg gcgaggagct
     2401 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     2461 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     2521 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct tcggctacgg
     2581 cctgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     2641 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     2701 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     2761 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     2821 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     2881 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     2941 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccgccct
     3001 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     3061 cgggatcact ctcggcatgg acgagctgta caagtaatag actagtcata tctagcgata
     3121 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     3181 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     3241 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     3301 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     3361 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3421 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3481 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     3541 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     3601 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     3661 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgctagaggg
     3721 ccctattcta tagtgtctct agagcggccg ccaccgcggg cccgggatct acgggtggca
     3781 tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac
     3841 cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa
     3901 tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag
     3961 ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc
     4021 aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt
     4081 ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac
     4141 catattggcc aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc
     4201 caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta
     4261 ggtaaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc
     4321 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     4381 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     4441 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     4501 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4561 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4621 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4681 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     4741 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4801 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     4861 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     4921 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     4981 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     5041 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     5101 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     5161 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     5221 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5281 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5341 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5401 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5461 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5521 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5581 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5641 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5701 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5761 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5821 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5881 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5941 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6001 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6061 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6121 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6181 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6241 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6301 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6361 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6421 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6481 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6541 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6601 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6661 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6721 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6781 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6841 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6901 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6961 ggaaaaacgc cagcaacgcg gccttttta
//
