Basic Vector Information
- Vector Name:
- pMT455
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7541 bp
- Type:
- Cloning vector
- Replication origin:
- R6K γ ori
- Source/Author:
- Taketani M, Zhang J, Zhang S, Triassi AJ
pMT455 vector Map
pMT455 vector Sequence
LOCUS 62056_17510 7541 bp DNA circular SYN 19-MAY-2020
DEFINITION Cloning vector pMT455, complete sequence.
ACCESSION MN991283
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7541)
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 7541)
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 7541)
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..7541
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(156..890)
/codon_start=1
/transl_table=11
/product="ErmG"
/label=ErmG
/protein_id="QJT41677.1"
/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"
protein_bind complement(3222..3240)
/label=tet operator
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 3284..3302
/label=tet operator
/note="bacterial operator O2 for the tetR and tetA genes"
regulatory 3303..3306
/label=rpiL*
/note="rpiL*"
/regulatory_class="terminator"
ncRNA 3307..3326
/label=sgRNA
/note="sgRNA-12"
/ncRNA_class="sgRNA"
misc_RNA 3327..3402
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
regulatory 3416..3467
/label=ECK120033736
/note="ECK120033736"
/regulatory_class="terminator"
regulatory 3476..3518
/label=cfxA
/note="cfxA"
/regulatory_class="promoter"
regulatory 3532..3535
/label=-33 sigma factor recognitions site
/note="-33 sigma factor recognitions site"
/regulatory_class="other"
regulatory 3543..3554
/label=PcfxA
/note="PcfxA"
/regulatory_class="promoter"
regulatory 3555..3562
/label=-7 sigma factor recognitions site
/note="-7 sigma factor recognitions site"
/regulatory_class="other"
ncRNA 3592..3611
/label=sgRNA
/note="sgRNA-12"
/ncRNA_class="sgRNA"
misc_feature 3612..3653
/label=chimera tracrRNA
/note="chimera tracrRNA"
regulatory 3658..3694
/label=derived from S. pyogenes
/note="derived from S. pyogenes"
/regulatory_class="terminator"
regulatory 3701..3752
/label=ECK120033736
/note="ECK120033736"
/regulatory_class="terminator"
regulatory 3761..3844
/label=PBF2884
/note="PBF2884"
/regulatory_class="promoter"
regulatory 3833..3836
/label=-33 sigma factor recognitions site
/note="-33 sigma factor recognitions site"
/regulatory_class="other"
misc_feature 3842..3843
/label=PAM
/note="PAM"
ncRNA 3845..3864
/label=sgRNA
/note="sgRNA-12"
/ncRNA_class="sgRNA"
misc_RNA 3870..3920
/label=sTRSV HHRz
/note="hammerhead ribozyme from the tobacco ringspot virus
satellite RNA (Khvorova et al., 2003)"
regulatory 3944..3986
/label=rpiL*
/note="rpiL*"
/regulatory_class="terminator"
CDS 3987..4499
/label=Nluc
/note="NanoLuc(R) luciferase"
regulatory 4510..4580
/label=L3S2P56
/note="L3S2P56"
/regulatory_class="terminator"
regulatory 4588..4870
/label=BT1311
/note="BT1311"
/regulatory_class="promoter"
regulatory 4831..4834
/label=-33 sigma factor recognitions site
/note="-33 sigma factor recognitions site"
/regulatory_class="other"
regulatory 4857..4864
/label=-7 sigma factor recognitions site
/note="-7 sigma factor recognitions site"
/regulatory_class="other"
regulatory 4871..4896
/label=BT1311
/note="BT1311"
/regulatory_class="ribosome_binding_site"
CDS 4897..5508
/codon_start=1
/transl_table=11
/product="TetR/AcrR family transcriptional regulator"
/label=TetR/AcrR family transcriptional regulator
/note="VFA0359-Bt"
/protein_id="QJT41680.1"
/translation="MQKKLTRSQQKHLDIINAAKEEFIEFGFLAANMDRITSSAEVSKR
TLYRHFESKEVLFESVLTIINDSVNESISYHFDPNKSTEEQLTEIAYKEIDVLYKTYGI
ALARTIVMEFLRQPEMAKTLIQNIYSIRAITQWFRSAIEAKRLKDADPKLMTDVYVSLF
QGLFFWPQVMHLDLEPHGEELSQKIETLTTIFLQSYGVAE"
regulatory 5525..5585
/label=L3S2P21
/note="L3S2P21"
/regulatory_class="terminator"
primer_bind complement(5711..5731)
/label=pNBU1_seq.REV
/note="pNBU1_seq.REV"
misc_recomb 5829..6158
/label=attN1 region
/note="attN1 region"
misc_feature 5956..5969
/label=CC Region
/note="CC Region"
misc_difference 5968
/label=mutated for higher integration efficiency
/note="mutated for higher integration efficiency"
CDS 6174..7511
/codon_start=1
/transl_table=11
/product="IntN1"
/label=IntN1
/note="NBU1 integrase"
/protein_id="QJT41681.1"
/translation="MKVTFIIKKAAKRYDTESMATIYVRFRNGRQLDSVAPTQLAINPN
LWDDKDECVKTKAVCNEEMRTHINEEIRQLKTYIEKVYQQEKEAIDKEWLKTTLDKFYH
PEKYFLPEEVVIKPTIGELFDEFLNKHPLSEVRKKNFRVVKRALLRYELYVRATKRGQK
GFILDVDLVTPDTLRDMWDFFQNEYQYYELYPSIYEAIPEKRTPQPRSKNTLIDCFSRI
RTFFLWCFDNKRTTNRPFDKFPIEECTYGTPYYITLEERDRIFNADLSATPQLAIQRDI
FIFQTLIGCRVSDLYRMTKLNVVNEAIEYIPKKTKEGNPVTVRVPLNDKAKEILERYKE
YEGKLLPFISEQKYNDAIKKIFKLAGVDRIVTILDPLTHNEIKRPIYEVASSHLARRTF
IGNIYKKVKDPNLVSALSGHKEGSKAFRRYRDIDEEMKKDLVKLLD"
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