LOCUS V032169 9510 bp DNA circular SYN 01-JAN-1980
DEFINITION synthetic circular DNA.
ACCESSION V032169
VERSION V032169
KEYWORDS .
SOURCE .
ORGANISM .
.
FEATURES Location/Qualifiers
promoter 1..204
/note="human cytomegalovirus (CMV) immediate early
promoter"
/label="CMV promoter"
CDS 293..1609
/label="TFAP2A(NM_001372066)"
/note="TFAP2A(NM_001372066)"
/gene="TFAP2A"
CDS 1616..1681
/codon_start=1
/product="three tandem FLAG® epitope tags, followed by an
enterokinase cleavage site"
/transl_table=1
/translation="DYKDHDGDYKDHDIDYKDDDDK"
/label="3xFLAG"
CDS 1691..2410
/codon_start=1
/note="mammalian codon-optimized"
/product="the original enhanced GFP (Yang et al., 1996)"
/transl_table=1
/translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
LLEFVTAAGITLGMDELYK*"
/label="EGFP"
promoter 2452..2663
/note="core promoter for human elongation factor
EF-1α"
/label="EF-1α core promoter"
LTR 2676..2944
/note="truncated 5' long terminal repeat (LTR) from human
T-cell leukemia virus (HTLV) type 1"
/label="5' LTR (truncated)"
CDS 2971..3570
/codon_start=1
/gene="pac from Streptomyces alboniger"
/note="confers resistance to puromycin"
/product="puromycin N-acetyltransferase"
/transl_table=1
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
/label="PuroR"
misc_feature 3577..4165
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
/label="WPRE"
LTR 4239..4472
/note="self-inactivating 3' long terminal repeat (LTR)
from HIV-1"
/label="3' LTR (ΔU3)"
polyA_signal 4544..4665
/note="SV40 polyadenylation signal"
/label="SV40 poly(A) signal"
rep_origin 4705..4840
/note="SV40 origin of replication"
/label="SV40 ori"
primer_bind complement(4873..4889)
/note="common sequencing primer, one of multiple similar
variants"
/label="M13 rev"
protein_bind 4897..4913
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG)."
/label="lac operator"
promoter complement(4921..4951)
/note="promoter for the E. coli lac operon"
/label="lac promoter"
protein_bind 4966..4987
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
/label="CAP binding site"
rep_origin complement(5275..5863)
/direction=LEFT
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
/label="ori"
CDS complement(6034..6894)
/codon_start=1
/gene="bla"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/product="β-lactamase"
/transl_table=1
/translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
SLIKHW*"
/label="AmpR"
promoter complement(6895..6999)
/gene="bla"
/label="AmpR promoter"
primer_bind 7473..7489
/note="common sequencing primer, one of multiple similar
variants"
/label="M13 fwd"
promoter 7504..7730
/note="Rous sarcoma virus enhancer/promoter"
/label="RSV promoter"
LTR 7731..7911
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
/label="5' LTR (truncated)"
misc_feature 7958..8083
/note="packaging signal of human immunodeficiency virus
type 1"
/label="HIV-1 Ψ"
misc_feature 8576..8809
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
/label="RRE"
CDS 8994..9038
/codon_start=1
/note="recognized by the 2H10 single-chain llama nanobody"
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/transl_table=1
/translation="KNEQELLELDKWASL"
/label="gp41 peptide"
misc_feature 9305..9421
/note="central polypurine tract and central termination
sequence of HIV-1"
/label="cPPT/CTS"