pMG_DV vector (V015655)

LOCUS       V015655                 6435 bp    DNA     circular SYN 23-OCT-2019
DEFINITION  Exported.
ACCESSION   V015655
VERSION     V015655
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6435)
  AUTHORS   Stach C, McCann M, O'Brien C, Le TS, Somia N, Chen X, Lee K, Fu HY,
            Daoutidis P, Zhao L, Hu WS, Smanski MJ.
  TITLE     Model-driven engineering of N-linked glycosylation in Chinese
            Hamster Ovary cells
  JOURNAL   ACS Synth Biol (2019) In press
   PUBMED   31596566
REFERENCE   2  (bases 1 to 6435)
  AUTHORS   Stach CS.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-AUG-2019) Biochemistry, Molecular Biology, and
            Biophysics; Chemical Engineering and Materials Science, University
            of Minnesota, 1479 Gortner Ave, Suite 344, Saint Paul, Minnesota
            55108, United States
REFERENCE   3  (bases 1 to 6435)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth
            Biol (2019) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (28-AUG-2019) Biochemistry, Molecular Biology, and Biophysics;
            Chemical Engineering and Materials Science, University of Minnesota,
            1479 Gortner Ave, Suite 344, Saint Paul, Minnesota 55108, United
            States"
FEATURES             Location/Qualifiers
     source          1..6435
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(21..54)
                     /label="FRT (minimal)"
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     misc_feature    55..65
                     /label="AarI site"
                     /note="AarI site"
     misc_feature    66..69
                     /label="Golden gate cloning scar"
                     /note="Golden gate cloning scar"
     misc_feature    70..77
                     /label="BbsI site"
                     /note="BbsI site"
     primer_bind     279..295
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    296..352
                     /label="MCS"
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(365..381)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(389..405)
                     /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)."
     promoter        complement(413..443)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(458..479)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    560..567
                     /label="BbsI site"
                     /note="BbsI site"
     misc_feature    568..571
                     /label="Golden gate cloning scar"
                     /note="Golden gate cloning scar"
     misc_feature    572..582
                     /label="AarI site"
                     /note="AarI site"
     promoter        588..1799
                     /label="UbC promoter"
                     /note="human ubiquitin C promoter"
     CDS             1831..2250
                     /gene="bsr"
                     /label="Blasticidin-S deaminase"
                     /note="Blasticidin-S deaminase from Bacillus cereus.
                     Accession#: P33967"
     polyA_signal    2257..2305
                     /label="poly(A) signal"
                     /note="synthetic polyadenylation signal"
     protein_bind    complement(2306..2339)
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2358..2396
                     /label="PhiC31 attP"
                     /note="PhiC31 attP"
     CDS             complement(2719..3279)
                     /codon_start=1
                     /transl_table=11
                     /product="kanamycin resistance protein"
                     /label="kanamycin resistance protein"
                     /protein_id="QFQ66234.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLHV
                     IYTLLLPRKYPSWLMQCGGCIRLIRLPAHSTTKRNIASSEHVLGWKPVLSIRMIWTKSI
                     RGSRQPNCSPGSRRACPTARISS"
     terminator      3812..3898
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3990..4017
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     rep_origin      4151..4739
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(4925..5065)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(6362..6379)
                     /label="6xHis"
                     /note="6xHis affinity tag"
     protein_bind    6400..6433
                     /label="attB"
                     /note="minimal attB site for the phi-C31 integrase (Groth
                     et al., 2000)"