Basic Vector Information
- Vector Name:
- pMT470
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9402 bp
- Type:
- Cloning vector
- Replication origin:
- R6K γ ori
- Source/Author:
- Taketani M, Zhang J, Zhang S, Triassi AJ
pMT470 vector Vector Map
pMT470 vector Sequence
LOCUS 62056_17530 9402 bp DNA circular SYN 19-MAY-2020
DEFINITION Cloning vector pMT470, complete sequence.
ACCESSION MN991287
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9402)
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 9402)
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 9402)
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..9402
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(156..890)
/codon_start=1
/transl_table=11
/product="ErmG"
/label=ErmG
/protein_id="QJT41696.1"
/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"
protein_bind 3288..3306
/label=tet operator
/note="bacterial operator O2 for the tetR and tetA genes"
misc_RNA 3331..3406
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
regulatory 3414..3474
/label=L3S3P41
/note="L3S3P41"
/regulatory_class="terminator"
misc_feature 3475..3478
/label=Bscar
/note="Bscar"
protein_bind complement(3567..3585)
/label=tet operator
/note="bacterial operator O2 for the tetR and tetA genes"
misc_feature 3652..3754
/label=M3
/note="M3"
regulatory 3755..3825
/label=ECK120034435
/note="ECK120034435"
/regulatory_class="terminator"
misc_feature 3826..3829
/label=Dscar
/note="Dscar"
regulatory 3830..3937
/label=PM1
/note="PM1"
/regulatory_class="promoter"
misc_feature 3938..4040
/label=M4
/note="M4"
regulatory 4041..4107
/label=ECK120033736
/note="ECK120033736"
/regulatory_class="terminator"
misc_feature 4108..4111
/label=Escar
/note="Escar"
regulatory 4112..4219
/label=PM1
/note="PM1"
/regulatory_class="promoter"
misc_feature 4220..4322
/label=M3
/note="M3"
regulatory 4323..4393
/label=ECK120034435
/note="ECK120034435"
/regulatory_class="terminator"
misc_feature 4394..4397
/label=Fscar
/note="Fscar"
regulatory 4398..4513
/label=P_BA
/note="P_BA"
/regulatory_class="promoter"
misc_feature 4514..4616
/label=M4
/note="M4"
regulatory 4617..4683
/label=ECK120033736
/note="ECK120033736"
/regulatory_class="terminator"
misc_feature 4684..4687
/label=scar
/note="nonstandard type: scar"
regulatory 4688..4803
/label=P_BA
/note="P_BA"
/regulatory_class="promoter"
misc_feature 4804..4906
/label=M1
/note="M1"
regulatory 4907..4967
/label=L3S3P41
/note="L3S3P41"
/regulatory_class="terminator"
misc_feature 4968..4971
/label=Cscar
/note="Cscar"
misc_feature 4972..4975
/label=Ascar
/note="Ascar"
regulatory 4976..5083
/label=PM4
/note="PM4"
/regulatory_class="promoter"
misc_RNA 5085..5135
/label=sTRSV HHRz
/note="hammerhead ribozyme from the tobacco ringspot virus
satellite RNA (Khvorova et al., 2003)"
regulatory 5159..5201
/label=rpiL*
/note="rpiL*"
/regulatory_class="terminator"
CDS 5202..5714
/label=Nluc
/note="NanoLuc(R) luciferase"
regulatory 5725..5795
/label=L3S2P56
/note="L3S2P56"
/regulatory_class="terminator"
misc_feature 5803..5806
/label=Bscar
/note="Bscar"
regulatory 5807..5914
/label=PM1
/note="PM1"
/regulatory_class="promoter"
misc_feature 5915..5989
/label=RiboJ00
/note="RiboJ00"
regulatory 5990..6032
/label=rpiL*
/note="rpiL*"
/regulatory_class="terminator"
misc_feature 6033..6038
/label=similar to luciferase reporter
/note="similar to luciferase reporter"
regulatory 6046..6116
/label=L3S2P56
/note="L3S2P56"
/regulatory_class="terminator"
misc_feature 6124..6127
/label=Dscar
/note="Dscar"
regulatory 6128..6235
/label=PM3
/note="PM3"
/regulatory_class="promoter"
misc_feature 6236..6310
/label=RiboJ00
/note="RiboJ00"
regulatory 6311..6353
/label=rpiL*
/note="rpiL*"
/regulatory_class="terminator"
misc_feature 6354..6359
/label=similar to luciferase reporter
/note="similar to luciferase reporter"
regulatory 6367..6437
/label=L3S2P56
/note="L3S2P56"
/regulatory_class="terminator"
misc_feature 6445..6448
/label=Cscar
/note="Cscar"
regulatory 6449..6731
/label=BT1311
/note="BT1311"
/regulatory_class="promoter"
regulatory 6692..6695
/label=-33 sigma factor recognitions site
/note="-33 sigma factor recognitions site"
/regulatory_class="other"
regulatory 6718..6725
/label=-7 sigma factor recognitions site
/note="-7 sigma factor recognitions site"
/regulatory_class="other"
CDS 6758..7369
/codon_start=1
/transl_table=11
/product="TetR/AcrR family transcriptional regulator"
/label=TetR/AcrR family transcriptional regulator
/note="VFA0359-Bt"
/protein_id="QJT41699.1"
/translation="MQKKLTRSQQKHLDIINAAKEEFIEFGFLAANMDRITSSAEVSKR
TLYRHFESKEVLFESVLTIINDSVNESISYHFDPNKSTEEQLTEIAYKEIDVLYKTYGI
ALARTIVMEFLRQPEMAKTLIQNIYSIRAITQWFRSAIEAKRLKDADPKLMTDVYVSLF
QGLFFWPQVMHLDLEPHGEELSQKIETLTTIFLQSYGVAE"
regulatory 7386..7446
/label=L3S2P21
/note="L3S2P21"
/regulatory_class="terminator"
primer_bind complement(7572..7592)
/label=pNBU1_seq.REV
/note="pNBU1_seq.REV"
misc_recomb 7690..8019
/label=attN1 region
/note="attN1 region"
misc_feature 7817..7830
/label=CC Region
/note="CC Region"
misc_difference 7829
/label=mutated for higher integration efficiency
/note="mutated for higher integration efficiency"
CDS 8035..9372
/codon_start=1
/transl_table=11
/product="IntN1"
/label=IntN1
/note="NBU1 integrase"
/protein_id="QJT41700.1"
/translation="MKVTFIIKKAAKRYDTESMATIYVRFRNGRQLDSVAPTQLAINPN
LWDDKDECVKTKAVCNEEMRTHINEEIRQLKTYIEKVYQQEKEAIDKEWLKTTLDKFYH
PEKYFLPEEVVIKPTIGELFDEFLNKHPLSEVRKKNFRVVKRALLRYELYVRATKRGQK
GFILDVDLVTPDTLRDMWDFFQNEYQYYELYPSIYEAIPEKRTPQPRSKNTLIDCFSRI
RTFFLWCFDNKRTTNRPFDKFPIEECTYGTPYYITLEERDRIFNADLSATPQLAIQRDI
FIFQTLIGCRVSDLYRMTKLNVVNEAIEYIPKKTKEGNPVTVRVPLNDKAKEILERYKE
YEGKLLPFISEQKYNDAIKKIFKLAGVDRIVTILDPLTHNEIKRPIYEVASSHLARRTF
IGNIYKKVKDPNLVSALSGHKEGSKAFRRYRDIDEEMKKDLVKLLD"
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