Basic Vector Information
- Vector Name:
- pMT448
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7194 bp
- Type:
- Cloning vector
- Replication origin:
- R6K γ ori
- Source/Author:
- Taketani M, Zhang J, Zhang S, Triassi AJ
pMT448 vector Map
pMT448 vector Sequence
LOCUS 62056_17490 7194 bp DNA circular SYN 19-MAY-2020
DEFINITION Cloning vector pMT448, complete sequence.
ACCESSION MN991279
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7194)
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 7194)
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 7194)
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..7194
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(156..890)
/codon_start=1
/transl_table=11
/product="ErmG"
/label=ErmG
/protein_id="QJT41657.1"
/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"
misc_feature 3123..3129
/label=Scar
/note="Scar"
regulatory 3134..3176
/label=cfxA
/note="cfxA"
/regulatory_class="promoter"
regulatory 3189..3192
/label=-33 sigma factor recognitions site
/note="-33 sigma factor recognitions site"
/regulatory_class="other"
misc_feature 3201..3212
/label=from PcfxA
/note="from PcfxA"
regulatory 3213..3220
/label=-7 sigma factor recognitions site
/note="-7 sigma factor recognitions site"
/regulatory_class="other"
ncRNA 3250..3269
/label=sgRNA
/note="sgRNA-7"
/ncRNA_class="sgRNA"
misc_RNA 3270..3345
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
regulatory 3353..3402
/label=L3S3P41
/note="L3S3P41"
/regulatory_class="terminator"
regulatory 3414..3489
/label=PBF2884
/note="PBF2884"
/regulatory_class="promoter"
regulatory 3486..3489
/label=-33 sigma factor recognitions site
/note="-33 sigma factor recognitions site"
/regulatory_class="other"
misc_feature 3495..3496
/label=PAM
/note="PAM"
ncRNA 3498..3517
/label=sgRNA
/note="sgRNA-7"
/ncRNA_class="sgRNA"
misc_RNA 3523..3573
/label=sTRSV HHRz
/note="hammerhead ribozyme from the tobacco ringspot virus
satellite RNA (Khvorova et al., 2003)"
regulatory 3597..3639
/label=rpiL*
/note="rpiL*"
/regulatory_class="terminator"
CDS 3640..4152
/label=Nluc
/note="NanoLuc(R) luciferase"
regulatory 4163..4233
/label=L3S2P56
/note="L3S2P56"
/regulatory_class="terminator"
regulatory 4241..4523
/label=BT1311
/note="BT1311"
/regulatory_class="promoter"
regulatory 4484..4487
/label=-33 sigma factor recognitions site
/note="-33 sigma factor recognitions site"
/regulatory_class="other"
regulatory 4510..4517
/label=-7 sigma factor recognitions site
/note="-7 sigma factor recognitions site"
/regulatory_class="other"
regulatory 4524..4549
/label=BT1311
/note="BT1311"
/regulatory_class="ribosome_binding_site"
CDS 4550..5161
/codon_start=1
/transl_table=11
/product="TetR/AcrR family transcriptional regulator"
/label=TetR/AcrR family transcriptional regulator
/note="VFA0359-Bt"
/protein_id="QJT41661.1"
/translation="MQKKLTRSQQKHLDIINAAKEEFIEFGFLAANMDRITSSAEVSKR
TLYRHFESKEVLFESVLTIINDSVNESISYHFDPNKSTEEQLTEIAYKEIDVLYKTYGI
ALARTIVMEFLRQPEMAKTLIQNIYSIRAITQWFRSAIEAKRLKDADPKLMTDVYVSLF
QGLFFWPQVMHLDLEPHGEELSQKIETLTTIFLQSYGVAE"
regulatory 5178..5238
/label=L3S2P21
/note="L3S2P21"
/regulatory_class="terminator"
primer_bind complement(5364..5384)
/label=pNBU1_seq.REV
/note="pNBU1_seq.REV"
misc_recomb 5482..5811
/label=attN1 region
/note="attN1 region"
misc_feature 5609..5622
/label=CC Region
/note="CC Region"
misc_difference 5621
/replace="g"
/label=mutated for higher integration efficiency
/note="mutated for higher integration efficiency"
CDS 5827..7164
/codon_start=1
/transl_table=11
/product="IntN1"
/label=IntN1
/note="NBU1 integrase"
/protein_id="QJT41660.1"
/translation="MKVTFIIKKAAKRYDTESMATIYVRFRNGRQLDSVAPTQLAINPN
LWDDKDECVKTKAVCNEEMRTHINEEIRQLKTYIEKVYQQEKEAIDKEWLKTTLDKFYH
PEKYFLPEEVVIKPTIGELFDEFLNKHPLSEVRKKNFRVVKRALLRYELYVRATKRGQK
GFILDVDLVTPDTLRDMWDFFQNEYQYYELYPSIYEAIPEKRTPQPRSKNTLIDCFSRI
RTFFLWCFDNKRTTNRPFDKFPIEECTYGTPYYITLEERDRIFNADLSATPQLAIQRDI
FIFQTLIGCRVSDLYRMTKLNVVNEAIEYIPKKTKEGNPVTVRVPLNDKAKEILERYKE
YEGKLLPFISEQKYNDAIKKIFKLAGVDRIVTILDPLTHNEIKRPIYEVASSHLARRTF
IGNIYKKVKDPNLVSALSGHKEGSKAFRRYRDIDEEMKKDLVKLLD"
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