LOCUS V029093 9984 bp DNA circular SYN 01-JAN-1980
DEFINITION synthetic circular DNA.
ACCESSION V029093
VERSION V029093
KEYWORDS .
SOURCE .
ORGANISM .
.
FEATURES Location/Qualifiers
enhancer 65..368
/note="human cytomegalovirus immediate early enhancer"
/label="CMV enhancer"
promoter 369..572
/note="human cytomegalovirus (CMV) immediate early
promoter"
/label="CMV promoter"
protein_bind 595..619
/bound_moiety="BP Clonase™"
/gene="mutant version of attB"
/note="recombination site for the Gateway® BP reaction"
/label="attB1"
CDS 619..2883
/label="SENP5(NM_152699)"
/note="SENP5(NM_152699)"
/gene="SENP5"
protein_bind complement(2882..2906)
/bound_moiety="BP Clonase™"
/gene="mutant version of attB"
/note="recombination site for the Gateway® BP reaction"
/label="attB2"
CDS 2908..2949
/codon_start=1
/product="epitope tag from simian virus 5"
/transl_table=1
/translation="GKPIPNPLLGLDST"
/label="V5 tag"
misc_feature 2991..3579
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
/label="WPRE"
LTR 3651..3884
/note="self-inactivating 3' long terminal repeat (LTR)
from HIV-1"
/label="3' LTR (ΔU3)"
polyA_signal 3962..4083
/note="SV40 polyadenylation signal"
/label="SV40 poly(A) signal"
rep_origin 4123..4258
/note="SV40 origin of replication"
/label="SV40 ori"
promoter complement(4279..4297)
/note="promoter for bacteriophage T7 RNA polymerase"
/label="T7 promoter"
primer_bind complement(4307..4323)
/note="common sequencing primer, one of multiple similar
variants"
/label="M13 fwd"
rep_origin 4465..4920
/direction=RIGHT
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
/label="f1 ori"
promoter 4946..5050
/gene="bla"
/label="AmpR promoter"
CDS 5051..5911
/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 6082..6670
/direction=RIGHT
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
/label="ori"
protein_bind 6958..6979
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
/label="CAP binding site"
promoter 6994..7024
/note="promoter for the E. coli lac operon"
/label="lac promoter"
protein_bind 7032..7048
/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 7056..7072
/note="common sequencing primer, one of multiple similar
variants"
/label="M13 rev"
promoter 7093..7111
/note="promoter for bacteriophage T3 RNA polymerase"
/label="T3 promoter"
promoter 7139..7365
/note="Rous sarcoma virus enhancer/promoter"
/label="RSV promoter"
LTR 7366..7546
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
/label="5' LTR (truncated)"
misc_feature 7593..7718
/note="packaging signal of human immunodeficiency virus
type 1"
/label="HIV-1 Ψ"
misc_feature 8211..8444
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
/label="RRE"
CDS 8629..8673
/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"
promoter 8848..9348
/note="human phosphoglycerate kinase 1 promoter"
/label="hPGK promoter"
CDS 9360..9755
/codon_start=1
/gene="Aspergillus terreus BSD"
/note="confers resistance to blasticidin"
/product="blasticidin S deaminase"
/transl_table=1
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
/label="BSD"
misc_feature 9815..9932
/note="central polypurine tract and central termination
sequence of HIV-1"
/label="cPPT/CTS"