LOCUS       Exported                4882 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHR-hSyn-EGFP-8X2C, complete sequence.
ACCESSION   MH458076
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4882)
  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 4882)
  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 4882)
  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..4882
                     /organism="Cloning vector pHR-hSyn-EGFP-8X2C"
                     /mol_type="other DNA"
                     /db_xref="taxon:2250251"
     misc_feature    1..635
                     /label=5LTR
                     /note="5LTR"
     LTR             1..634
                     /label=3 LTR
                     /note="3 long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1535
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2049..2166
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     regulatory      2227..2707
                     /regulatory_class="promoter"
                     /note="hSyn"
     promoter        2254..2701
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      2714..2722
                     /regulatory_class="ribosome_binding_site"
     regulatory      2717..2726
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2723..3442
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             2726..3442
                     /codon_start=1
                     /product="EGFP"
                     /label=EGFP
                     /protein_id="AWX65704.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     ncRNA           3457..3892
                     /label=miRNA
                     /note="microRNA"
                     /ncRNA_class="miRNA"
     misc_feature    3942..4542
                     /label=WPRE
                     /note="WPRE"
     misc_feature    3952..4540
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(4423..4434)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4543..4559)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    4647..4882
                     /label=3LTR
                     /note="3LTR"