LOCUS       Exported                4259 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector AAV-CamKIIalpha-ChR2-EGFP, complete sequence.
ACCESSION   MH458082
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4259)
  AUTHORS   Tseng H-a., Ta T, Gritton HJ, Man H-Y.
  TITLE     A microRNA-based gene targeting tool for virally labeling 
            interneurons in the rodent cortex
  JOURNAL   Cell Rep (2018) In press
REFERENCE   2  (bases 1 to 4259)
  AUTHORS   Keaveney MK, Han X.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-JUN-2018) Biomedical Engineering, Boston University, 
            610 Commonwealth Ave, Boston, MA 02215, USA
REFERENCE   3  (bases 1 to 4259)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: APE Plasmid Editor v. v2.0.53.c
            Sequencing Technology :: Illumina
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4259
                     /organism="Cloning vector AAV-CamKIIalpha-ChR2-EGFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:2250245"
     repeat_region   1..101
                     /label=5ITR
                     /note="5ITR"
     regulatory      197..1487
                     /regulatory_class="promoter"
                     /note="CamKIIalpha"
     regulatory      1493..1504
                     /regulatory_class="ribosome_binding_site"
     regulatory      1499..1508
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1505..2431
                     /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"
     CDS             1508..3169
                     /codon_start=1
                     /product="ChR2-EGFP"
                     /label=ChR2-EGFP
                     /protein_id="AWX65710.1"
                     /translation="DYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGTN
                     GAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKNP
                     SMLYLATGHRVQWLRYAEWLLTCPVILIHLSNLTGLSNDYSRRTMGLLVSDIGTIVWGA
                     TSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVSW
                     GMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDIR
                     KTTKLNIGGTEIEVETLVEDEAEAGAVPAAPVVAVSKGEELFTGVVPILVELDGDVNGH
                     KFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFF
                     KSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEY
                     NYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLS
                     TQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     CDS             2453..3169
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     misc_feature    3209..3797
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     misc_feature    3211..3798
                     /label=WPRE
                     /note="WPRE"
     CDS             complement(3680..3691)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3800..3816)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    3846..3967
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   4070..4259
                     /label=3ITR
                     /note="3ITR"