LOCUS       sequence-126565-        6600 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of ChR2-Venus for neuronal excitation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6600)
  AUTHORS   
  TITLE     The subcellular organization of neocortical excitatory connections.
  JOURNAL   Nature. 2009 Jan 18.
  PUBMED    19151697
REFERENCE   2  (bases 1 to 6600)
  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: "Nature. 
            2009 Jan 18."
COMMENT     Sequence Label: pACAGW-ChR2-Venus-AAV
FEATURES             Location/Qualifiers
     source          1..6600
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   56..185
                     /label=AAV2 ITR
     polyA_signal    347..468
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(384..403)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     438..457
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    511..1099
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             617..628
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1026..1046
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     complement(1153..1172)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          1155..1287
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     CDS             complement(join(1313..2026,2027..2029,2030..2032))
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1358..1379)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1707..1726
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1966..1987
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2029..2038
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(2066..2992)
                     /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"
     primer_bind     complement(3043..3062)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3124..3146)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3425..3702)
                     /label=chicken beta-actin promoter
     repeat_region   4013..4142
                     /label=AAV2 ITR
     rep_origin      4214..4669
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4301..4320)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4511..4532
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4695..4799
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4800..4868,4869..5660)
                     /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(5018..5037)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5831..6419
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6320..6339
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg
       61 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
      121 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
      181 ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa
      241 gatccttatc gattttacca catttgtaga ggttttactt gctttaaaaa acctcccaca
      301 tctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
      361 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
      421 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgctc
      481 gaagcggccg cccgggtcga cggtatcgat gcggggaggc ggcccaaagg gagatccgac
      541 tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg
      601 aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa
      661 tccaggtggc aacacaggcg agcagccaag gaaaggacga tgatttcccc gacaacacca
      721 cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga
      781 tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg
      841 tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac
      901 gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg
      961 aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt
     1021 atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt
     1081 tgtaatccag aggttgatta tcgataagct tgatatcgaa ttcgaatcga ctagtcgcga
     1141 gctccactcg acacctgtgg agagaaaggc aaagtggatg tcagtaagac caataggtgc
     1201 ctatcagaaa cgcaagagtc ttctctgtct cgacaagccc agtttctatt ggtctcctta
     1261 aacctgtctt gtaaccttga tacttacctg cggcctcgag tccggggccg ctttacttgt
     1321 acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt
     1381 gatcgcgctt ctcgttgggg tctttgctca gcttggactg gtagctcagg tagtggttgt
     1441 cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct
     1501 gcacgccgcc gtcctcgatg ttgtggcgga tcttgaagtt ggccttgatg ccgttcttct
     1561 gcttgtcggc ggtgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca
     1621 ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt
     1681 cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg
     1741 tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt
     1801 ggtcggggta gcgggcgaag cactgcaggc cgtagcccag ggtggtcacg agggtgggcc
     1861 agggcacggg cagcttgccg gtggtgcaga tcagcttcag ggtcagcttg ccgtaggtgg
     1921 catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct
     1981 cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggtggcga
     2041 ccggtggctt gccggtgccc ttgttgaccg cgccagcctc ggcctcgtcc tccaccagcg
     2101 tctcgacctc aatctcagtg ccaccaatgt tcaatttggt ggtcttgcga atgtcgccgt
     2161 ggatgaggat atgctcgtgg atcagcacgc gcaggtagtg gccgagcaga ccccagcagt
     2221 tcttcgacat caggtcaatg atggtgtggc cgacggtgga gccgtacacg ctcaggacgc
     2281 cgaagccctc ggggccgagg atgaacagga tggggaacat accccatgat acgaagaaga
     2341 gccaagccat gccagtcacc acctggcgac accggccctt cggcacggtg tggtaaccct
     2401 cgatgtaggc cttggcagcg tgaaagaacg tgttagcacc ataacacaga cccaggcaga
     2461 agaagatgac cttgacgtat ccggtggcca tggcggaagt ggcgccccac acaattgtgc
     2521 caatatcaga cacaagcaga cccatggtgc gcctgctgta gtcgttggac aagcccgtca
     2581 ggtttgacag gtgaatgaga atgaccgggc aggtgagaag ccactcggcg taacgcaacc
     2641 actggacgcg gtggcctgtg gctagataca gcatggacgg gttcttaaac tcgaagaaga
     2701 actcgagaat caccttgacc atctcgatag cgcacacata gatctcctcc cagccgcagg
     2761 ttgacttcca tgtttggtag gcgtaaaaca taagcagtag gatggagaag ccagcagcaa
     2821 gccattgcag cacgttcgac gccgtttggg caccgtttgt gccacgcgac tcaatccagc
     2881 ccgcgcagta acactggtcc tcaggcacaa gtacagagcc attgacgact actgggttcg
     2941 ttacaaatag cagctcgcgc ccaacggcac tcagggcgcc tccataatcc atgctagcac
     3001 cggtggatcc cgatccaatt ctttgccaaa atgatgagac agcacaataa ccagcacgtt
     3061 gcccaggagc tgtaggaaaa agaagaaggc atgaacatgg ttagcagagg ctctagagcc
     3121 gccggtcaca cgccagaagc cgaaccccgc cctgccccgt cccccccgaa ggcagccgtc
     3181 cccccgcgga cagccccgag ccgctccccc cgcacaaagg gccctcccgg agcccctcaa
     3241 ggctttcacg cagccacaga aaagaaacaa gccgtcatta aaccaagcgc taattacagc
     3301 ccggaggaga agggccgtcc cgcccgctca cctgtgggag taacgcggtc agtcagagcc
     3361 ggggcgggcg gcgcgaggcg gcggcggagc ggggcacggg gcgaaggcag cgcgcagcga
     3421 ctcccgcccg ccgcgcgctt cgctttttat agggccgccg ccgccgccgc ctcgccataa
     3481 aaggaaactt tcggagcgcg ccgctctgat tggctgccgc cgcacctctc cgcctcgccc
     3541 cgccccgccc ctcgccccgc cccgccccgc ctggcgcgcg cccccccccc ccccccgccc
     3601 ccatcgctgc acaaaataat taaaaaataa ataaatacaa aattgggggt ggggaggggg
     3661 gggagatggg gagagtgaag cagaacgtgg ggctcacctc gaccatggta atagcgatga
     3721 ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat
     3781 gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca
     3841 cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga
     3901 aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat ggtactcttc
     3961 ctagagcatg gctacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc
     4021 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     4081 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     4141 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     4201 aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4261 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     4321 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4381 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     4441 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4501 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4561 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     4621 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
     4681 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4741 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4801 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4861 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4921 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4981 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5041 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5101 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5161 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5221 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5281 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5341 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5401 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5461 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5521 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5581 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5641 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5701 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5761 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5821 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5881 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5941 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6001 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6061 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6121 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6181 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6241 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6301 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6361 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6421 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6481 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     6541 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
//
