pMT470 vector (V015764)

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"