Basic Vector Information
- Vector Name:
- pTXB1-Tn5
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7716 bp
- Type:
- Bacterial Expression
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- T7 promoter
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- T7 promoter primer
- 3' Primer:
- GATTGCCATGCCGGTCAAGG
pTXB1-Tn5 vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pTXB1-Tn5 vector Sequence
LOCUS 40924_44549 7716 bp DNA circular SYN 13-MAY-2021
DEFINITION IPTG-inducible expression of Transposon Tn5 fused to Mxe Intein and
Chitin-binding domain..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7716)
AUTHORS Picelli S, Bjorklund AK, Reinius B, Sagasser S, Winberg G, Sandberg
R
TITLE Tn5 transposase and tagmentation procedures for massively-scaled
sequencing projects.
JOURNAL Genome Res. 2014 Jul 30. pii: gr.177881.114.
PUBMED 25079858
REFERENCE 2 (bases 1 to 7716)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 7716)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Genome Res.
2014 Jul 30. pii: gr.177881.114."
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7716
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 126..148
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(145..333)
/codon_start=1
/label=rop
/note="Rop protein, which maintains plasmids at low copy
number"
/translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
protein_bind complement(856..877)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(893..1972)
/codon_start=1
/label=lacI
/note="lac repressor"
/translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
promoter complement(1973..2050)
/label=lacI promoter
terminator 2203..2289
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 2386..2472
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator complement(2572..2615)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
promoter 2790..2808
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 2809..2833
/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)."
CDS 2877..4304
/codon_start=1
/label=Tn5 transposase
/note="transposase from the bacterial Tn5 transposon
(Reznikoff, 1993)"
/translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS
ITISSEGSKAMQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR
HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG
KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGL
YLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA
VLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER
QRMEEPDNLERMVSILSFVAVRLLQLRESFTPPQALRAQGLLKEAEHVESQSAETVLTP
DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS
KLDGFLAAKDLMAQGIKI"
CDS 4305..4898
/codon_start=1
/label=Mxe GyrA intein
/note="modified GyrA intein (Southworth et al., 1999)"
/translation="CITGDALVALPEGESVRIADIVPGARPNSDNAIDLKVLDRHGNPV
LADRLFHSGEHPVYTVRTVEGLRVTGTANHPLLCLVDVAGVPTLLWKLIDEIKPGDYAV
IQRSAFSVDCAGFARGKPEFAPTTYTVGVPGLVRFLEAHHRDPDAQAIADELTDGRFYY
AKVASVTDAGVQPVYSLRVDTADHAFITNGFVSHA"
CDS 4929..5084
/codon_start=1
/label=CBD
/note="chitin binding domain from chitinase A1 (Watanabe et
al., 1994)"
/translation="TTNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNV
PALWQLQ"
terminator 5163..5210
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter 5270..5374
/label=AmpR promoter
CDS 5375..6232
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin complement(6277..6790)
/direction=LEFT
/label=M13 ori
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
rep_origin 6901..7489
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7643..7660
/label=L4440
/note="L4440 vector, forward primer"
primer_bind complement(7682..7701)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
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