pGEX_LacI_TetR_chPylRS_IPYE_MjTyrRS vector (V015840)

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LOCUS       V015840                 7190 bp    DNA     circular SYN 19-APR-2020
DEFINITION  Exported.
ACCESSION   V015840
VERSION     V015840
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7190)
  AUTHORS   Fischer EC, Hashimoto K, Zhang Y, Feldman AW, Dien VT, Karadeema RJ,
            Adhikary R, Ledbetter MP, Krishnamurthy R, Romesberg FE.
  TITLE     New codons for efficient production of unnatural proteins in a
            semi-synthetic organism
  JOURNAL   Nat. Chem. Biol. (2020) In press
REFERENCE   2  (bases 1 to 7190)
  AUTHORS   Fischer EC.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-DEC-2019) Department of Chemistry, The Scripps
            Research Institute, 10550 North Torrey Pines Road, La Jolla, CA
            92037, USA
REFERENCE   3  (bases 1 to 7190)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. Chem.
            Biol. (2020) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (27-DEC-2019) Department of Chemistry, The Scripps Research
            Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA"
FEATURES             Location/Qualifiers
     source          1..7190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      19..607
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     regulatory      717..786
                     /label="AmpR promoter"
                     /note="AmpR promoter"
                     /regulatory_class="promoter"
     CDS             846..1466
                     /label="TetR"
                     /note="tetracycline repressor TetR"
     terminator      complement(1491..1577)
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     promoter        1893..1970
                     /label="lacIq promoter"
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             1971..3050
                     /label="lacI"
                     /note="lac repressor"
     promoter        3171..3201
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    3209..3225
                     /label="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             3254..4171
                     /gene="tyrS"
                     /label="Tyrosine--tRNA ligase"
                     /note="Tyrosine--tRNA ligase from Methanocaldococcus
                     jannaschii (strain ATCC 43067 / DSM 2661 / JAL-1 / JCM
                     10045 / NBRC 100440). Accession#: Q57834"
     promoter        4371..4399
                     /label="tac promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    4407..4423
                     /label="lacI binding site"
                     /bound_moiety="lacI"
                     /note="lac repressor binds to the lac operator to inhibit
                     transcription in E. coli; lac operator"
     CDS             4446..5705
                     /codon_start=1
                     /transl_table=11
                     /product="chimeric PylRS variant"
                     /label="chimeric PylRS variant"
                     /note="chimeric PylRS variant; Methanosarcina mazei/barkeri
                     chPylRS(IPYE)"
                     /protein_id="QIZ64222.1"
                     /translation="MDKKPLDVLISATGLWMSRTGTLHKIKHYEISRSKIYIEMACGDH
                     LVVNNSRSCRPARAFRYHKYRKTCKRCRVSDEDINNFLTRSTEGKTSVKVKVVSEPKVK
                     KAMPKSVSRAPKPLENPVSAKASTDTSRSVPSPAKSTPNSPVPTSASAPALTKSQTDRL
                     EVLLNPKDEISLNSGKPFRELESELLSRRKKDLQQIYAEERENYLGKLEREITRFFVDR
                     GFLEIKSPILIPLEYIERMGIDNDTELSKQIFRVDKNFCLRPMLAPNLYNYLRKLDRAL
                     PDPIKIFEIGPCYRKESDGKEHLEEFTMLNFCQMGSGCTRENLESIITDFLNHLGIDFK
                     IVGDSCMVYGDTLDVMHGDLELSSAVVGPIPLDREWGIDKPWIGAGFGLERLLKVKHDF
                     KNIKRAARSESYYNGISTNL"
     terminator      5929..5976
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     regulatory      6073..6177
                     /label="AmpR promoter"
                     /note="AmpR promoter"
                     /regulatory_class="promoter"
     CDS             6178..7035
                     /label="AmpR"
                     /note="beta-lactamase"