Basic Vector Information
- Vector Name:
- pT-GEM(2)
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8124 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Diao F, Ironfield H, Luan H, Diao F, Shropshire WC, Ewer J, Marr E, Potter CJ, Landgraf M, White BH.
pT-GEM(2) vector Map
pT-GEM(2) vector Sequence
LOCUS 40924_42024 8124 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pT-GEM(2), complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8124)
AUTHORS Diao F, Ironfield H, Luan H, Diao F, Shropshire WC, Ewer J, Marr E,
Potter CJ, Landgraf M, White BH.
TITLE Plug-and-Play Genetic Access to Drosophila Cell Types using
Exchangeable Exon Cassettes
JOURNAL Cell Rep 10 (8), 1410-1421 (2015)
PUBMED 25732830
REFERENCE 2 (bases 1 to 8124)
AUTHORS Diao F, Luan H, White BH.
TITLE Direct Submission
JOURNAL Submitted (22-JAN-2015) Laboratory of Molecular Biology, NIMH, NIH,
35 Convent Dr, Bethesda, MD 20850, USA
REFERENCE 3 (bases 1 to 8124)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 8124)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell Rep";
date: "2015"; volume: "10"; issue: "8"; pages: "1410-1421"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(22-JAN-2015) Laboratory of Molecular Biology, NIMH, NIH, 35 Convent
Dr, Bethesda, MD 20850, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Assembly Method :: Vector NTI v. Version 11
Sequencing Technology :: Illumina
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..8124
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 3..19
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 26..44
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 52..71
/label=MCS
/note="MCS"
protein_bind 72..171
/label=phage phi-C31 attP
/note="attachment site of phage phi-C31"
misc_feature 178..277
/label=MHC splice acceptor site
/note="MHC splice acceptor site"
misc_feature 278..311
/label=linker
/note="linker"
CDS 318..371
/codon_start=1
/label=T2A
/note="2A peptide from Thosea asigna virus capsid protein"
/translation="EGRGSLLTCGDVEENPGP"
CDS 372..3014
/codon_start=1
/label=GAL4
/note="GAL4 transcriptional activator"
/translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVSIDSAAHHDNSTIPLDF
MPRDALHGFDWSEEDDMSDGLPFLKTDPNNNGFFGDGSLLCILRSIGFKPENYTNSNVN
RLPTMITDRYTLASRSTTSRLLQSYLNNFHPYCPIVHSPTLMMLYNNQIEIASKDQWQI
LFNCILAIGAWCIEGESTDIDVFYYQNAKSHLTSKVFESGSIILVTALHLLSRYTQWRQ
KTNTSYNFHSFSIRMAISLGLNRDLPSSFSDSSILEQRRRIWWSVYSWEIQLSLLYGRS
IQLSQNTISFPSSVDDVQRTTTGPTIYHGIIETARLLQVFTKIYELDKTVTAEKSPICA
KKCLMICNEIEEVSRQAPKFLQMDISTTALTNLLKEHPWLSFTRFELKWKQLSLIIYVL
RDFFTNFTQKKSQLEQDQNDHQSYEVKRCSIMLSDAAQRTVMSVSSYMDNHNVTPYFAW
NCSYYLFNAVLVPIKTLLSNSKSNAENNETAQLLQQINTVLMLLKKLATFKIQTCEKYI
QVLEEVCAPFLLSQCAIPLPHISYNNSNGSAIKNIVGSATIAQYPTLPEENVNNISVKY
VSPGSVGPSPVPLKSGASFSDLVKLLSNRPPSRNSPVTIPRSTPSHRSVTPFLGQQQQL
QSLVPLTPSALFGGANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
EDTPPNPKKE"
misc_recomb 3521..3554
/label=loxP
/note="loxP"
protein_bind 3521..3554
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
CDS 3786..4460
/codon_start=1
/label=DsRed1
/note="wild-type DsRed"
/translation="MRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRHH
LFL"
intron 4540..4605
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 4735..4755
/codon_start=1
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/translation="PKKKRKV"
polyA_signal 5027..5161
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
protein_bind complement(5168..5201)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
protein_bind complement(5208..5307)
/label=phage phi-C31 attP
/note="attachment site of phage phi-C31"
misc_feature 5306..5325
/label=MCS
/note="MCS"
promoter complement(5338..5356)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(5377..5393)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(5401..5417)
/label=lac operator
/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(5425..5455)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(5470..5491)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5779..6367)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6541..7398)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(7399..7503)
/label=AmpR promoter
rep_origin 7530..7985
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
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