pTHSSe_71 vector (V002648)

LOCUS       V002648                10275 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V002648
VERSION     V002648
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10275)
  AUTHORS   Segall-Shapiro TH, Sontag ED, Voigt CA.
  TITLE     Engineered promoters enable constant gene expression at any copy
            number in bacteria
  JOURNAL   Nat. Biotechnol. (2018) In press
   PUBMED   29553576
REFERENCE   2  (bases 1 to 10275)
  AUTHORS   Segall-Shapiro TH, Sontag ED, Voigt CA.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-MAR-2018) Synthetic Biology Center, Department of
            Biological Engineering, Massachusetts Institute of Technology, 77
            Massachusetts Avenue, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 10275)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10275)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat.
            Biotechnol. (2018) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (06-MAR-2018) Synthetic Biology Center, Department of Biological
            Engineering, Massachusetts Institute of Technology, 77 Massachusetts
            Avenue, Cambridge, MA 02139, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10275
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    9..12
                     /label="GGAG seam"
                     /note="GGAG seam"
     regulatory      13..73
                     /label="L3S2P21"
                     /note="L3S2P21"
                     /regulatory_class="terminator"
     regulatory      78..146
                     /label="PT7A1w3"
                     /note="PT7A1w3"
                     /regulatory_class="promoter"
     ncRNA           147..225
                     /product="SarJ"
                     /label="ribozyme"
                     /note="insulator"
                     /ncRNA_class="ribozyme"
     regulatory      226..245
                     /label="RBSsp2"
                     /note="RBSsp2"
                     /regulatory_class="ribosome_binding_site"
     CDS             246..2885
                     /codon_start=1
                     /product="TALEsp2"
                     /label="TALEsp2"
                     /protein_id="AVR55202.1"
                     /translation="MVDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVAL
                     SQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQL
                     DTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPDQVVAIASHDGGKQALETVQRLL
                     PVLCQDHGLTPEQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQ
                     ALETVQRLLPVLCQAHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQDHGLTPDQVVA
                     IASNGGGKQALETVQRLLPVLCQDHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAH
                     GLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNNGGKQALETVQR
                     LLPVLCQDHGLTPDQVVAIASNIGGKQALETVQRLLPVLCQDHGLTPEQVVAIASHDGG
                     KQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQV
                     VAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNIGGKQALETVQRLLPVLCQ
                     DHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETV
                     QRLLPVLCQAHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQDHGLTPEQVVAIASNG
                     GGRPALESIVAQLSRPDPALAALTNDHLVALACLGGRPALDAVKKGLPHAPALIKRTNR
                     RIPERTSHRVADHAQVVRVLGFFQCHSHPAQAFDDAMTQFGMSRHGLLQLFRRVGVTEL
                     EARSGTLPPASQRWDRILQASGMKRAKPSPTSTQTPDQASLHAFADSLERDLDAPSPMH
                     EGDQTRAS"
     regulatory      complement(2891..2980)
                     /label="ECK120029600"
                     /note="ECK120029600"
                     /regulatory_class="terminator"
     regulatory      2985..3059
                     /label="PUPsp2"
                     /note="PUPsp2"
                     /regulatory_class="promoter"
     misc_feature    3010..3016
                     /label="bfpT7A1"
                     /note="bfpT7A1"
     protein_bind    complement(3018..3035)
                     /label="TALEsp2 binding site"
                     /bound_moiety="TALEsp2"
     misc_RNA        3061..3111
                     /label="sTRSV HHRz"
                     /note="hammerhead ribozyme from the tobacco ringspot virus
                     satellite RNA (Khvorova et al., 2003)"
     regulatory      3135..3157
                     /label="RBSVioEs"
                     /note="RBSVioEs"
                     /regulatory_class="ribosome_binding_site"
     CDS             3158..3733
                     /codon_start=1
                     /product="VioE"
                     /label="VioE"
                     /protein_id="AVR55201.1"
                     /translation="MENREPPLLPARWSSAYVSYWSPMLPDDQLTSGYCWFDYERDICR
                     IDGLFNPWSERDTGYRLWMSEVGNAASGRTWKQKVAYGRERTALGEQLCERPLDDETGP
                     FAELFLPRDVLRRLGARHIGRRVVLGREADGWRYQRPGKGPSTLYLDAASGTPLRMVTG
                     DEASRASLRDFPNVSEAEIPDAVFAAKR"
     regulatory      3734..3757
                     /label="RBSVioAs"
                     /note="RBSVioAs"
                     /regulatory_class="ribosome_binding_site"
     CDS             3758..5011
                     /gene="vioA"
                     /label="Flavin-dependent L-tryptophan oxidase VioA"
                     /note="Flavin-dependent L-tryptophan oxidase VioA from
                     Chromobacterium violaceum (strain ATCC 12472 / DSM 30191 /
                     JCM 1249 / NBRC 12614 / NCIMB 9131 / NCTC 9757).
                     Accession#: Q9S3V1"
     regulatory      5015..5038
                     /label="RBSVioBs"
                     /note="RBSVioBs"
                     /regulatory_class="ribosome_binding_site"
     CDS             5039..8032
                     /gene="vioB"
                     /label="2-imino-3-(indol-3-yl)propanoate dimerase"
                     /note="2-imino-3-(indol-3-yl)propanoate dimerase from
                     Chromobacterium violaceum (strain ATCC 12472 / DSM 30191 /
                     JCM 1249 / NBRC 12614 / NCIMB 9131 / NCTC 9757).
                     Accession#: Q9S3V0"
     terminator      8050..8121
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      8137..8164
                     /label="T7Te terminator"
                     /note="phage T7 early transcription terminator"
     misc_feature    8183..8203
                     /label="BioBrick suffix"
                     /note="universal suffix for all parts"
     terminator      8204..8261
                     /label="his operon terminator"
                     /note="This putative transcriptin terminator from the E.
                     coli his operon has a 2-bp deletion introduced during
                     synthesis. Its efficiency has not been determined."
     rep_origin      8433..9021
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(9222..10013)
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     terminator      complement(10208..10251)
                     /label="bacterial terminator"
                     /note="putative bacterial transcription terminator"
     misc_feature    10254..10275
                     /label="BioBrick prefix"
                     /note="BioBrick prefix for parts that do not start with
                     'ATG'"