Basic Vector Information
- Vector Name:
- pMT462
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9657 bp
- Type:
- Cloning vector
- Replication origin:
- R6K γ ori
- Source/Author:
- Taketani M, Zhang J, Zhang S, Triassi AJ
pMT462 vector Vector Map
pMT462 vector Sequence
LOCUS 62056_17515 9657 bp DNA circular SYN 19-MAY-2020
DEFINITION Cloning vector pMT462, complete sequence.
ACCESSION MN991284
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9657)
AUTHORS Taketani M, Zhang J, Zhang S, Triassi AJ, Huang YJ, Griffith LG,
Voigt CA.
TITLE Genetic circuit design automation for the gut resident species
Bacteroides thetaiotaomicron
JOURNAL Nat. Biotechnol. (2020) In press
PUBMED 32231334
REFERENCE 2 (bases 1 to 9657)
AUTHORS Taketani M, Voigt CA.
TITLE Direct Submission
JOURNAL Submitted (25-JAN-2020) Biological Engineering, MIT, Synthetic
Biology Center 500 Technology Square NE47-140, Cambridge, MA 02139,
USA
REFERENCE 3 (bases 1 to 9657)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat.
Biotechnol. (2020) In press"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(25-JAN-2020) Biological Engineering, MIT, Synthetic Biology Center
500 Technology Square NE47-140, Cambridge, MA 02139, USA"
FEATURES Location/Qualifiers
source 1..9657
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(156..890)
/codon_start=1
/transl_table=11
/product="ErmG"
/label=ErmG
/protein_id="QJT41682.1"
/translation="MNKVNIKDSQNFITSKYHIEKIMNCISLDEKDNIFEIGAGKGHFT
AGLVKRCNFVTAIEIDSKLCEVTRNKLLNYPNYQIVNDDILKFTFPSHNPYKIFGSIPY
NISTNIIRKIVFESSATISYLIVEYGFAKMLLDTNRSLALLLMAEVDISILAKIPRYYF
HPKPKVDSTLIVLKRKPAKMAFKERKKYETFVMKWVNKEYEKLFTKNQFNKALKHARIY
DINNISFEQFVSLFNSYKIFNG"
CDS 1378..2235
/label=AmpR
/note="beta-lactamase"
oriT complement(2369..2478)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
rep_origin 2649..3037
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
regulatory 3078..3107
/label=tL17
/note="tL17"
/regulatory_class="terminator"
regulatory 3123..3130
/label=L3S3P21
/note="L3S3P21"
/regulatory_class="terminator"
misc_feature 3134..3137
/label=Ascar
/note="Ascar"
regulatory 3138..3245
/label=PM1
/note="PM1"
/regulatory_class="promoter"
misc_RNA 3266..3341
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
regulatory 3349..3452
/label=ECK120029600
/note="ECK120029600"
/regulatory_class="terminator"
misc_difference 3360
/replace="c"
misc_feature 3453..3456
/label=Bscar
/note="Bscar"
protein_bind 3607..3625
/label=tet operator
/note="bacterial operator O2 for the tetR and tetA genes"
misc_feature 3630..3732
/label=M4
/note="M4"
regulatory 3733..3799
/label=ECK120033736
/note="ECK120033736"
/regulatory_class="terminator"
misc_feature 3800..3803
/label=Dscar
/note="Dscar"
regulatory 3804..3919
/label=P_BA
/note="P_BA"
/regulatory_class="promoter"
misc_feature 3920..4022
/label=M2
/note="M2"
regulatory 4023..4126
/label=ECK120029600
/note="ECK120029600"
/regulatory_class="terminator"
misc_feature 4127..4130
/label=Escar
/note="Escar"
regulatory 4131..4238
/label=PM1
/note="PM1"
/regulatory_class="promoter"
misc_feature 4239..4341
/label=M4
/note="M4"
regulatory 4342..4408
/label=ECK120033736
/note="ECK120033736"
/regulatory_class="terminator"
misc_feature 4409..4412
/label=Fscar
/note="Fscar"
regulatory 4413..4528
/label=P_BA
/note="P_BA"
/regulatory_class="promoter"
misc_feature 4529..4631
/label=M1
/note="M1"
regulatory 4632..4692
/label=L3S3P41
/note="L3S3P41"
/regulatory_class="terminator"
misc_feature 4693..4696
/label=scar
/note="nonstandard type: scar"
protein_bind complement(4785..4803)
/label=tet operator
/note="bacterial operator O2 for the tetR and tetA genes"
misc_feature 4870..4972
/label=M1
/note="M1"
regulatory 4973..5033
/label=L3S3P41
/note="L3S3P41"
/regulatory_class="terminator"
misc_feature 5034..5037
/label=Cscar
/note="Cscar"
misc_feature 5038..5041
/label=Ascar
/note="Ascar"
regulatory 5042..5149
/label=PM2
/note="PM2"
/regulatory_class="promoter"
misc_RNA 5151..5201
/label=sTRSV HHRz
/note="hammerhead ribozyme from the tobacco ringspot virus
satellite RNA (Khvorova et al., 2003)"
regulatory 5225..5267
/label=rpiL*
/note="rpiL*"
/regulatory_class="terminator"
CDS 5268..5780
/label=Nluc
/note="NanoLuc(R) luciferase"
regulatory 5791..5861
/label=L3S2P56
/note="L3S2P56"
/regulatory_class="terminator"
misc_feature 5869..5872
/label=Bscar
/note="Bscar"
regulatory 5873..5980
/label=PM4
/note="PM4"
/regulatory_class="promoter"
misc_feature 5981..6055
/label=RiboJ00
/note="RiboJ00"
CDS 6099..6611
/label=Nluc
/note="NanoLuc(R) luciferase"
regulatory 6622..6692
/label=L3S2P56
/note="L3S2P56"
/regulatory_class="terminator"
misc_feature 6700..6703
/label=Cscar
/note="Cscar"
regulatory 6704..6986
/label=BT1311
/note="BT1311"
/regulatory_class="promoter"
regulatory 6947..6950
/label=-33 sigma factor recognitions site
/note="-33 sigma factor recognitions site"
/regulatory_class="other"
regulatory 6973..6980
/label=-7 sigma factor recognitions site
/note="-7 sigma factor recognitions site"
/regulatory_class="other"
regulatory 6987..7012
/label=BT1311
/note="BT1311"
/regulatory_class="ribosome_binding_site"
CDS 7013..7624
/codon_start=1
/transl_table=11
/product="TetR/AcrR family transcriptional regulator"
/label=TetR/AcrR family transcriptional regulator
/note="VFA0359-Bt"
/protein_id="QJT41686.1"
/translation="MQKKLTRSQQKHLDIINAAKEEFIEFGFLAANMDRITSSAEVSKR
TLYRHFESKEVLFESVLTIINDSVNESISYHFDPNKSTEEQLTEIAYKEIDVLYKTYGI
ALARTIVMEFLRQPEMAKTLIQNIYSIRAITQWFRSAIEAKRLKDADPKLMTDVYVSLF
QGLFFWPQVMHLDLEPHGEELSQKIETLTTIFLQSYGVAE"
regulatory 7641..7701
/label=L3S2P21
/note="L3S2P21"
/regulatory_class="terminator"
primer_bind complement(7827..7847)
/label=pNBU1_seq.REV
/note="pNBU1_seq.REV"
misc_recomb 7945..8274
/label=attN1 region
/note="attN1 region"
misc_feature 8072..8085
/label=CC Region
/note="CC Region"
misc_difference 8084
/label=mutated for higher integration efficiency
/note="mutated for higher integration efficiency"
CDS 8290..9627
/codon_start=1
/transl_table=11
/product="IntN1"
/label=IntN1
/note="NBU1 integrase"
/protein_id="QJT41687.1"
/translation="MKVTFIIKKAAKRYDTESMATIYVRFRNGRQLDSVAPTQLAINPN
LWDDKDECVKTKAVCNEEMRTHINEEIRQLKTYIEKVYQQEKEAIDKEWLKTTLDKFYH
PEKYFLPEEVVIKPTIGELFDEFLNKHPLSEVRKKNFRVVKRALLRYELYVRATKRGQK
GFILDVDLVTPDTLRDMWDFFQNEYQYYELYPSIYEAIPEKRTPQPRSKNTLIDCFSRI
RTFFLWCFDNKRTTNRPFDKFPIEECTYGTPYYITLEERDRIFNADLSATPQLAIQRDI
FIFQTLIGCRVSDLYRMTKLNVVNEAIEYIPKKTKEGNPVTVRVPLNDKAKEILERYKE
YEGKLLPFISEQKYNDAIKKIFKLAGVDRIVTILDPLTHNEIKRPIYEVASSHLARRTF
IGNIYKKVKDPNLVSALSGHKEGSKAFRRYRDIDEEMKKDLVKLLD"
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