Basic Vector Information
- Vector Name:
- pMT449
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7237 bp
- Type:
- Cloning vector
- Replication origin:
- R6K γ ori
- Source/Author:
- Taketani M, Zhang J, Zhang S, Triassi AJ
pMT449 vector Vector Map
pMT449 vector Sequence
LOCUS 62056_17495 7237 bp DNA circular SYN 19-MAY-2020
DEFINITION Cloning vector pMT449, complete sequence.
ACCESSION MN991280
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7237)
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 7237)
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 7237)
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..7237
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(156..890)
/codon_start=1
/transl_table=11
/product="ErmG"
/label=ErmG
/protein_id="QJT41662.1"
/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"
misc_difference 3062
/replace="g"
regulatory 3078..3107
/label=tL17
/note="tL17"
/regulatory_class="terminator"
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"
regulatory 3201..3212
/label=PcfxA
/note="PcfxA"
/regulatory_class="promoter"
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-8"
/ncRNA_class="sgRNA"
misc_RNA 3270..3345
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
regulatory 3353..3448
/label=ECK120029600
/note="ECK120029600"
/regulatory_class="terminator"
regulatory 3449..3456
/label=ECK120029600
/note="ECK120029600"
/regulatory_class="terminator"
regulatory 3457..3540
/label=PBF2884
/note="PBF2884"
/regulatory_class="promoter"
regulatory 3529..3532
/label=-33 sigma factor recognitions site
/note="-33 sigma factor recognitions site"
/regulatory_class="other"
misc_feature 3538..3539
/label=PAM
/note="PAM"
ncRNA 3541..3560
/label=sgRNA
/note="sgRNA-8"
/ncRNA_class="sgRNA"
misc_RNA 3566..3616
/label=sTRSV HHRz
/note="hammerhead ribozyme from the tobacco ringspot virus
satellite RNA (Khvorova et al., 2003)"
regulatory 3640..3682
/label=rpiL*
/note="rpiL*"
/regulatory_class="terminator"
CDS 3683..4195
/label=Nluc
/note="NanoLuc(R) luciferase"
regulatory 4206..4276
/label=L3S2P56
/note="L3S2P56"
/regulatory_class="terminator"
regulatory 4284..4566
/label=BT1311
/note="BT1311"
/regulatory_class="promoter"
regulatory 4527..4530
/label=-33 sigma factor recognitions site
/note="-33 sigma factor recognitions site"
/regulatory_class="other"
regulatory 4553..4560
/label=-7 sigma factor recognitions site
/note="-7 sigma factor recognitions site"
/regulatory_class="other"
regulatory 4567..4592
/label=BT1311
/note="BT1311"
/regulatory_class="ribosome_binding_site"
CDS 4593..5204
/codon_start=1
/transl_table=11
/product="TetR/AcrR family transcriptional regulator"
/label=TetR/AcrR family transcriptional regulator
/note="VFA0359-Bt"
/protein_id="QJT41665.1"
/translation="MQKKLTRSQQKHLDIINAAKEEFIEFGFLAANMDRITSSAEVSKR
TLYRHFESKEVLFESVLTIINDSVNESISYHFDPNKSTEEQLTEIAYKEIDVLYKTYGI
ALARTIVMEFLRQPEMAKTLIQNIYSIRAITQWFRSAIEAKRLKDADPKLMTDVYVSLF
QGLFFWPQVMHLDLEPHGEELSQKIETLTTIFLQSYGVAE"
regulatory 5221..5281
/label=L3S2P21
/note="L3S2P21"
/regulatory_class="terminator"
misc_recomb 5525..5854
/label=attN1 region
/note="attN1 region"
misc_feature 5652..5665
/label=CC Region
/note="CC Region"
misc_difference 5664
/replace="g"
/label=mutated for higher integration efficiency
/note="mutated for higher integration efficiency"
CDS 5870..7207
/codon_start=1
/transl_table=11
/product="IntN1"
/label=IntN1
/note="NBU1 integrase"
/protein_id="QJT41666.1"
/translation="MKVTFIIKKAAKRYDTESMATIYVRFRNGRQLDSVAPTQLAINPN
LWDDKDECVKTKAVCNEEMRTHINEEIRQLKTYIEKVYQQEKEAIDKEWLKTTLDKFYH
PEKYFLPEEVVIKPTIGELFDEFLNKHPLSEVRKKNFRVVKRALLRYELYVRATKRGQK
GFILDVDLVTPDTLRDMWDFFQNEYQYYELYPSIYEAIPEKRTPQPRSKNTLIDCFSRI
RTFFLWCFDNKRTTNRPFDKFPIEECTYGTPYYITLEERDRIFNADLSATPQLAIQRDI
FIFQTLIGCRVSDLYRMTKLNVVNEAIEYIPKKTKEGNPVTVRVPLNDKAKEILERYKE
YEGKLLPFISEQKYNDAIKKIFKLAGVDRIVTILDPLTHNEIKRPIYEVASSHLARRTF
IGNIYKKVKDPNLVSALSGHKEGSKAFRRYRDIDEEMKKDLVKLLD"
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