LOCUS Exported 10960 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Translation reporter.
ACCESSION .
VERSION .
KEYWORDS pcDNA4TO-24xGCN4_v4-kif18b-24xPP7
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10960)
AUTHORS Yan X, Hoek TA, Vale RD, Tanenbaum ME
TITLE Dynamics of Translation of Single mRNA Molecules In Vivo.
JOURNAL Cell. 2016 May 5;165(4):976-89. doi: 10.1016/j.cell.2016.04.034.
PUBMED 27153498
REFERENCE 2 (bases 1 to 10960)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.cell.2016.04"; journalName: "Cell"; date: "2016-05-5- 5";
volume: "165"; issue: "4"; pages: "976-89"
FEATURES Location/Qualifiers
source 1..10960
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
protein_bind 820..838
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 841..859
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 869..893
/label=LNCX
/note="Human CMV promoter, forward primer"
regulatory 984..993
/regulatory_class="other"
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1059..1115
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1131..1187
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1203..1259
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1275..1331
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1347..1403
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1419..1475
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1491..1547
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1563..1619
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1635..1691
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1707..1763
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1779..1835
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1851..1907
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1923..1979
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1995..2051
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 2067..2123
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 2139..2195
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 2211..2267
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 2283..2339
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 2355..2411
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 2427..2483
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 2499..2555
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 2571..2627
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 2715..2771
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
primer_bind complement(6971..6988)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 6977..7201
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 7247..7675
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7334..7353)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7544..7565
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(7684..7704)
/label=pBABE 3
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 7689..8018
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 7869..8004
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 7931..7950
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 8066..8113
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 8132..8506
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
polyA_signal 8636..8757
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(8673..8692)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 8727..8746
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(8806..8822)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8806..8822)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(8819..8841)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 8830..8846
/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)."
promoter complement(8854..8884)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8899..8920
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(9037..9054)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(9208..9793)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(9288..9307)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(9964..10824)
/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 10587..10606
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(10825..10929)
/gene="bla"
/label=AmpR promoter
