Basic Vector Information
- Vector Name:
- pMK228
- Antibiotic Resistance:
- Streptomycin
- Length:
- 4588 bp
- Type:
- Cloning vector
- Replication origin:
- p15A ori
- Source/Author:
- Hartsough LA, Kotlajich MV, Han B, Lin C-C.J.
pMK228 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 ℃.
pMK228 vector Sequence
LOCUS 62056_17020 4588 bp DNA circular SYN 27-JUL-2020
DEFINITION Cloning vector pMK228, complete sequence.
ACCESSION MN617162
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4588)
AUTHORS Hartsough LA, Kotlajich MV, Han B, Lin C-C.J., Gambill L, Wang MC,
Tabor JJ.
TITLE Optogenetic control of gut bacterial metabolism
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 4588)
AUTHORS Hartsough LA, Kotlajich MV, Han B, Lin C-C.J., Gambill L, Wang MC,
Tabor JJ.
TITLE Direct Submission
JOURNAL Submitted (26-OCT-2019) Bioengineering, Rice University, 6500 Main
St., Houston, TX 77030, USA
REFERENCE 3 (bases 1 to 4588)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(26-OCT-2019) Bioengineering, Rice University, 6500 Main St.,
Houston, TX 77030, USA"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..4588
/mol_type="other DNA"
/organism="synthetic DNA construct"
regulatory 24..171
/note="specR promoter; from KD13"
/regulatory_class="promoter"
CDS 172..960
/label=SmR
/note="aminoglycoside adenylyltransferase (Murphy, 1985)"
terminator 975..1069
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
rep_origin 1183..1728
/direction=RIGHT
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin."
misc_feature 1984..2018
/label=constitutive promoter J23106
/note="constitutive promoter J23106"
misc_feature 2019..2053
/label=B0034
/note="B0034"
CDS 2054..4315
/codon_start=1
/transl_table=11
/product="CcaS"
/label=CcaS
/protein_id="QLL27773.1"
/translation="MGKFLIPIEFVFLAIAMTCYLWHRQNQERRRIEISIKQQTQRERF
INQITQHIRQSLNLETVLNTTVAEVKTLLQVDRVLIYRIWQDGTGSAITESVNANYPSI
LGRTFSDEVFPVEYHQAYTKGKVRAINDIDQDDIEICLADFVKQFGVKSKLVVPILQHN
RASSLDNESEFPYLWGLLITHQCAFTRPWQPWEVELMKQLANQVAIAIQQSELYEQLQQ
LNKDLENRVEKRTQQLAATNQSLRMEISERQKTEAALRHTNHTLQSLIAASPRGIFTLN
LADQIQIWNPTAERIFGWTETEIIAHPELLTSNILLEDYQQFKQKVLSGMVSPSLELKC
QKKDGSWIEIVLSAAPLLDSEENIAGLVAVVADITEQKRQAEQIRLLQSVVVNTNDAVV
ITEAEPIDDPGPRILYVNEAFTKITGYTAEEMLGKTPRVLQGPKTSRTELDRVRQAISQ
WQSVTVEVINYRKDGSEFWVEFSLVPVANKTGFYTHWIAVQRDVTERRRTEEVRLALER
EKELSRLKTRFFSMASHEFRTPLSTALAAAQLLENSEVAWLDPDKRSRNLHRIQNSVKN
MVQLLDDILIINRAEAGKLEFNPNWLDLKLLFQQFIEEIQLSVSDQYYFDFICSAQDTK
ALVDERLVRSILSNLLSNAIKYSPGGGQIKIALSLDSEQIIFEVTDQGIGISPEDQKQI
FEPFHRGKNVRNITGTGLGLMVAKKCVDLHSGSILLKSAVDQGTTVTICLKRYNHLPRA
"
terminator 4336..4407
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 4423..4450
/label=T7Te terminator
/note="phage T7 early transcription terminator"
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