pOtta_sfGFP__pOsym_mKate2 vector (V016136)

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LOCUS       62056_18310        5113 bp DNA     circular SYN 16-AUG-2021
DEFINITION  Cloning vector pOtta_sfGFP__pOsym_mKate2, complete sequence.
ACCESSION   MZ198787
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5113)
  AUTHORS   Groseclose TM, Hersey AN, Huang BD, Wilson CJ.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   2  (bases 1 to 5113)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (13-MAY-2021) Chemical "
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5113
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(400..494)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(528..1319)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     regulatory      1506..1534
                     /label=trc promoter
                     /note="trc promoter"
                     /regulatory_class="promoter"
     regulatory      1506..1511
                     /label=trc promoter -35 hexamer
                     /note="trc promoter -35 hexamer"
                     /regulatory_class="promoter"
     regulatory      1512..1528
                     /label=operator Otta
                     /note="operator Otta"
                     /regulatory_class="other"
     regulatory      1529..1534
                     /label=trc promoter -10 hexamer
                     /note="trc promoter -10 hexamer"
                     /regulatory_class="promoter"
     regulatory      1553..1564
                     /label=strong bacterial RBS
                     /note="strong bacterial RBS"
                     /regulatory_class="ribosome_binding_site"
     CDS             1571..2284
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Nager et al., 2011)"
     terminator      2299..2385
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(2926..3873)
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
     rep_origin      complement(3921..4143)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     protein_bind    4256..4275
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     regulatory      4275..4280
                     /label=trc promoter -10 hexamer
                     /note="trc promoter -10 hexamer"
                     /regulatory_class="promoter"
     regulatory      4299..4310
                     /label=strong bacterial RBS
                     /note="strong bacterial RBS"
                     /regulatory_class="ribosome_binding_site"
     CDS             4317..5012
                     /label=mKate2
                     /note="monomeric far-red fluorescent protein (Shcherbo et
                     al., 2009)"
     terminator      complement(5030..5073)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"