Basic Vector Information
- Vector Name:
- pNM58
- Antibiotic Resistance:
- Tetracycline
- Length:
- 9993 bp
- Type:
- Cloning vector
- Replication origin:
- p15A ori
- Source/Author:
- Gonzalez LM, Mukhitov N, Voigt CA.
pNM58 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 ℃.
pNM58 vector Sequence
LOCUS 62056_18100 9993 bp DNA circular SYN 16-DEC-2019
DEFINITION Cloning vector pNM58, complete sequence.
ACCESSION MN443783
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9993)
AUTHORS Gonzalez LM, Mukhitov N, Voigt CA.
TITLE Resilient living materials built by printing bacterial spores
JOURNAL Nat. Chem. Biol. (2019) In press
PUBMED 31792444
REFERENCE 2 (bases 1 to 9993)
AUTHORS Mukhitov N, Gonzalez LM, Voigt CA.
TITLE Direct Submission
JOURNAL Submitted (11-SEP-2019) Biological Engineering, MIT, NE47-140, 500
Tech Square, Cambridge, MA 02139, USA
REFERENCE 3 (bases 1 to 9993)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat. Chem.
Biol. (2019) In press"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(11-SEP-2019) Biological Engineering, MIT, NE47-140, 500 Tech
Square, Cambridge, MA 02139, USA"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..9993
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 251..795
/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."
CDS 1643..2350
/codon_start=1
/label=yeGFP
/note="yeast-enhanced green fluorescent protein"
/translation="KGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFI
CTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNY
KTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNF
KIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLEFV
TAAGITHGMDELYK"
terminator 2383..2430
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(6308..7165)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHF"
promoter complement(7166..7270)
/label=AmpR promoter
CDS complement(7437..8810)
/codon_start=1
/label=TcR
/note="tetracycline efflux protein"
/translation="VNTSYSQSNLRHNQILIWLCILSFFSVLNEMVLNVSLPDIANDFN
KPPASTNWVNTAFMLTFSIGTAVYGKLSDQLGIKRLLLFGIIINCFGSVIGFVGHSFFS
LLIMARFIQGAGAAAFPALVMVVVARYIPKENRGKAFGLIGSIVAMGEGVGPAIGGMIA
HYIHWSYLLLIPMITIITVPFLMKLLKKEVRIKGHFDIKGIILMSVGIVFFMLFTTSYS
ISFLIVSVLSFLIFVKHIRKVTDPFVDPGLGKNIPFMIGVLCGGIIFGTVAGFVSMVPY
MMKDVHQLSTAEIGSVIIFPGTMSVIIFGYIGGILVDRRGPLYVLNIGVTFLSVSFLTA
SFLLETTSWFMTIIIVFVLGGLSFTKTVISTIVSSSLKQQEAGAGMSLLNFTSFLSEGT
GIAIVGGLLSIPLLDQRLLPMEVDQSTYLYSNLLLLFSGIIVISWLVTLNVYKHSQRDF
"
CDS complement(join(9013..9993,1..21))
/codon_start=1
/label=repB
/note="RepB replication protein"
/translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
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