LOCUS       sequence-47435-e        6564 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Viral expression of ArchT-GFP for optogenetic inhibition.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6564)
  AUTHORS   Han X, Chow BY, Zhou H, Klapoetke NC, Chuong A, Rajimehr R, Yang A, 
            Baratta MV, Winkle J, Desimone R, Boyden ES
  TITLE     A high-light sensitivity optical neural silencer: development and 
            application to optogenetic control of non-human primate cortex.
  JOURNAL   Front Syst Neurosci. 2011;5:18. Epub 2011 Apr 13.
  PUBMED    21811444
REFERENCE   2  (bases 1 to 6564)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Front Syst 
            Neurosci."; date: "2011"; volume: "5"; pages: "18. Epub 2011 Apr 13"
COMMENT     Sequence Label: pAAV-CamKII-ArchT-GFP
FEATURES             Location/Qualifiers
     source          1..6564
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1932..2387)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2069..2090)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2281..2300
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2459..2588
                     /label=AAV2 ITR
     promoter        2661..3950
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      3960..3969
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4725..4727,4728..5441)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4725..4727,4728..5438)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(4767..4788)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5028..5047)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5375..5396
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5466..6054
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5519..5539)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5937..5948)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6097..6218
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6108..6127)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6162..6181
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   6380..6509
                     /label=AAV2 ITR
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg
     2521 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
     2581 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag
     2641 gaagatcgta ggttaattaa cattatggcc ttaggtcact tcatctccat ggggttcttc
     2701 ttctgatttt ctagaaaatg agatgggggt gcagagagct tcctcagtga cctgcccagg
     2761 gtcacatcag aaatgtcaga gctagaactt gaactcagat tactaatctt aaattccatg
     2821 ccttgggggc atgcaagtac gatatacaga aggagtgaac tcattagggc agatgaccaa
     2881 tgagtttagg aaagaagagt ccagggcagg gtacatctac accacccgcc cagccctggg
     2941 tgagtccagc cacgttcacc tcattatagt tgcctctctc cagtcctacc ttgacgggaa
     3001 gcacaagcag aaactgggac aggagcccca ggagaccaaa tcttcatggt ccctctggga
     3061 ggatgggtgg ggagagctgt ggcagaggcc tcaggagggg ccctgctgct cagtggtgac
     3121 agataggggt gagaaagcag acagagtcat tccgtcagca ttctgggtct gtttggtact
     3181 tcttctcacg ctaaggtggc ggtgtgatat gcacaatggc taaaaagcag ggagagctgg
     3241 aaagaaacaa ggacagagac agaggccaag tcaaccagac caattcccag aggaagcaaa
     3301 gaaaccatta cagagactac aagggggaag ggaaggagag atgaattagc ttcccctgta
     3361 aaccttagaa cccagctgtt gccagggcaa cggggcaata cctgtctctt cagaggagat
     3421 gaagttgcca gggtaactac atcctgtctt tctcaaggac catcccagaa tgtggcaccc
     3481 actagccgtt accatagcaa ctgcctcttt gccccactta atcccatccc gtctgttaaa
     3541 agggccctat agttggaggt gggggaggta ggaagagcga tgatcacttg tggactaagt
     3601 ttgttcacat ccccttctcc aaccccctca gtacatcacc ctgggagaac aaggtccact
     3661 tgcttctggg cccacacagt cctgcagtat tgtgtatata aggccagggc aacggaggag
     3721 caggttttga agtgaaaggc aggcaggtgt tggggaggca gttaccgggg caacgggaac
     3781 agggcgtttc ggaggtggtt gccatgggga cctggatgct gacgaaggct cgcgaggctg
     3841 tgagcagcca cagtgccctg ctcagaagcc ccaagctcgt caatcaagct ggttctccat
     3901 ttgcactcag gagcacgggc aggcgagtgg cccctagttc tgggggcagc gggggatccg
     3961 ccaccatgga ccccatcgct ctgcaggcag gatacgacct gctgggcgac ggaaggccag
     4021 agaccctgtg gctgggaatc ggaaccctgc tgatgctgat cggcaccttc tacttcatcg
     4081 tgaagggctg gggcgtgacc gacaaggagg ccagggagta ctacagcatc acaatcctgg
     4141 tgcccggcat cgccagcgcc gcctacctga gcatgttctt cggcatcggc ctgaccgagg
     4201 tgaccgtggc cggcgaggtg ctggacatct actacgccag atacgccgac tggctgttca
     4261 ccaccccttt gcttctgctc gacctggccc tgctggctaa ggtggacagg gtgagcatcg
     4321 gaaccctggt gggagtggac gccctgatga tcgtgaccgg cctgatcggc gccctgagcc
     4381 acaccccact ggctaggtac agctggtggc tgttcagcac catctgcatg atcgtggtgc
     4441 tgtacttcct ggctaccagc ctgagggctg ctgctaagga gaggggacca gaggtggcta
     4501 gcaccttcaa caccctgacc gccctggtgc tggtgctgtg gaccgcctac cccatcctgt
     4561 ggatcatcgg aaccgaggga gctggagtgg tgggactggg aatcgagacc ctgctgttca
     4621 tggtgctgga cgtgaccgcc aaagtgggct tcggcttcat cctgctgagg agcagggcca
     4681 tcctgggcga caccgaggcc cccgagcccc gaccggtagt agcagtgagc aagggcgagg
     4741 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     4801 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     4861 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     4921 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     4981 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     5041 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     5101 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     5161 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     5221 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     5281 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     5341 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     5401 ccgccgggat cactctcggc atggacgagc tgtacaagta agaattcgat atcaagctta
     5461 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     5521 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     5581 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     5641 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     5701 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     5761 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     5821 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     5881 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     5941 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     6001 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     6061 accgtcgacc cgggcggccg cttcgagcag acatgataag atacattgat gagtttggac
     6121 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     6181 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     6241 ttatgtttca ggttcagggg gagatgtggg aggtttttta aagcaagtaa aacctctaca
     6301 aatgtggtaa aatcgataag gatcttccta gagcatggct acgtagataa gtagcatggc
     6361 gggttaatca ttaactacaa ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     6421 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     6481 gcggcctcag tgagcgagcg agcgcgcagc tgcattaatg aatcggccaa cgcgcgggga
     6541 gaggcggttt gcgtattggg cgct
//
