| 载体基本信息 |
| 载体名称 |
pCCLc-MND-A0201-SABR-Backbone |
载体抗性 |
Ampicillin |
| 载体长度 |
10927 bp |
载体类型 |
Mammalian Expression, Lentiviral |
| 筛选标记 |
surface myc-tag |
拷贝数 |
High Copy |
| 启动子 |
MND |
5引物 |
GCTCCCCGAGCTCAATAAAAG |
| 3引物 |
TGGCTAAGATCTACAGCTGCCTT |
pCCLc-MND-A0201-SABR-Backbone 载体载体图谱
pCCLc-MND-A0201-SABR-Backbone 载体载体序列
LOCUS Exported 10927 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION 3rd gen transfer vector. HLA-A0201 linked to b2-microglobulin and
CD28-CD3z signaling domain. Contains a stuffer fragment flanked by
BsmBI sites at the epitope cloning site..
ACCESSION .
VERSION .
KEYWORDS pCCLc-MND-A0201-SABR-Backbone
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10927)
AUTHORS Joglekar AV, Leonard MT, Jeppson JD, Swift M, Li G, Wong S, Peng S,
TITLE T cell antigen discovery via signaling and antigen-presenting
bifunctional receptors
JOURNAL Nature Methods (2019) volume 16, pages 191–198.
REFERENCE 2 (bases 1 to 10927)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nature
Methods (2019) volume 16, pages 191–198."
FEATURES Location/Qualifiers
source 1..10927
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 1..22
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 37..67
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 75..91
/label=lac operator
/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-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 80..102
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 99..115
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 99..115
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 134..154
/label=T3
/note="T3 promoter, forward primer"
promoter 136..154
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
enhancer 255..634
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 635..838
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 789..809
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 853..1033
/label=5 LTR (truncated)
/note="truncated 5 long terminal repeat (LTR) from HIV-1"
misc_feature 1080..1205
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1698..1931
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 2116..2160
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 2457..2574
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
CDS 7065..7130
/codon_start=1
/product="2A peptide from foot-and-mouth disease virus
polyprotein"
/label=F2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="VKQTLNFDLLKLAGDVESNPGP"
CDS 7215..7244
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
LTR 7627..7860
/label=3 LTR (Delta-U3)
/note="self-inactivating 3 long terminal repeat (LTR) from
HIV-1"
polyA_signal 7932..8053
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(7969..7988)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 8023..8042
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 8093..8228
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 8155..8174
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(8248..8267)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(8249..8267)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(8277..8294)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(8277..8293)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8286..8308)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 8435..8890
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(8522..8541)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 8732..8753
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 8916..9020
/gene="bla"
/label=AmpR promoter
CDS 9021..9881
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(9239..9258)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 10052..10640
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 10541..10560
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 10794..10811
/label=L4440
/note="L4440 vector, forward primer"