LOCUS V034955 11003 bp DNA circular SYN 01-JAN-1980
DEFINITION synthetic circular DNA.
ACCESSION V034955
VERSION V034955
KEYWORDS .
SOURCE .
ORGANISM .
.
FEATURES Location/Qualifiers
polyA_signal 216..350
/note="SV40 polyadenylation signal"
/label="SV40 poly(A) signal"
promoter 398..502
/gene="bla"
/label="AmpR promoter"
CDS 503..1363
/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"
rep_origin 1534..2119
/direction=RIGHT
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
/label="ori"
protein_bind 2407..2428
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
/label="CAP binding site"
promoter 2443..2473
/note="promoter for the E. coli lac operon"
/label="lac promoter"
protein_bind 2481..2497
/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"
primer_bind 2505..2521
/note="common sequencing primer, one of multiple similar
variants"
/label="M13 rev"
LTR 2650..3283
/note="3' long terminal repeat (LTR) from HIV-1"
/label="3' LTR"
misc_feature 3490..4078
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
/label="WPRE"
CDS complement(4092..4691)
/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"
promoter complement(4700..5029)
/note="SV40 enhancer and early promoter"
/label="SV40 promoter"
CDS complement(5040..5786)
/codon_start=1
/product="modified rtTA protein that binds tightly to
promoters containing the tet operator in the presence of
doxycycline"
/transl_table=1
/translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
DALDDFDLDMLPADALDDFDLDMLPG*"
/label="Tet-On® 3G"
promoter complement(5805..6315)
/note="human phosphoglycerate kinase 1 promoter"
/label="hPGK promoter"
promoter 6332..6699
/note="3rd-generation Tet-responsive promoter that can be
activated by binding of Tet-On® 3G, modified to eliminate
binding sites for endogenous mammalian transcription
factors"
/label="TRE3GS promoter"
CDS 6712..8385
/label="GLUD1(NM_005271)"
/note="GLUD1(NM_005271)"
/gene="GLUD1"
CDS 8392..8457
/codon_start=1
/product="three tandem FLAG® epitope tags, followed by an
enterokinase cleavage site"
/transl_table=1
/translation="DYKDHDGDYKDHDIDYKDDDDK"
/label="3xFLAG"
CDS 8467..8496
/codon_start=1
/product="10xHis affinity tag"
/transl_table=1
/translation="HHHHHHHHHH"
/label="10xHis"
polyA_signal 8683..8817
/note="SV40 polyadenylation signal"
/label="SV40 poly(A) signal"
misc_feature 8861..8976
/note="central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)"
/label="cPPT/CTS"
CDS complement(9239..9283)
/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 9468..9701
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
/label="RRE"
misc_feature 10198..10323
/note="packaging signal of human immunodeficiency virus
type 1"
/label="HIV-1 Ψ"
LTR 10370..11003
/note="3' long terminal repeat (LTR) from HIV-1"
/label="3' LTR"