pB-tetO2-mTET3cd-mCherry vector (V016489)

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LOCUS       62056_3395        7514 bp DNA     circular SYN 27-APR-2021
DEFINITION  Cloning vector pB-tetO2-mTET3cd-mCherry, complete sequence.
ACCESSION   MW139646
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7514)
  AUTHORS   Hore TA.
  TITLE     Pronounced specificity of the TET enzyme catalytic domain guides 
            cellular function
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7514)
  AUTHORS   Hore TA.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-OCT-2020) Department of Anatomy, University of Otago, 
            270 Great King Street, Dunedin, Otago 9016, New Zealand
REFERENCE   3  (bases 1 to 7514)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (20-OCT-2020) Department of Anatomy, University of Otago, 270 Great 
            King Street, Dunedin, Otago 9016, New Zealand"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: Geneious v. March 2019
            Assembly Name         :: pB-tetO2-mTET3cd-mCherry
            Coverage              :: 463
            Sequencing Technology :: MiSeq
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7514
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     598..614
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        621..639
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   702..936
                     /label=LTR
                     /rpt_family="PiggyBAC LTR"
                     /note="LTR"
     protein_bind    1140..1158
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      join(1177..1195,1214..1232,1251..1269,1288..1306,1325..1343,
                     1362..1380,1399..1417)
                     /label=tetO2
                     /note="tetO2"
                     /regulatory_class="response_element"
     protein_bind    1543..1567
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1573..2922
                     /codon_start=1
                     /transl_table=11
                     /product="mTET3cd"
                     /label=mTET3cd
                     /protein_id="QQM12951.1"
                     /translation="MRPLEFPTCDCVEQIVEKDEGPYYTHLGSGPTVASIRELMEDRYG
                     EKGKAIRIEKVIYTGKEGKSSRGCPIAKWVIRRHTLEEKLLCLVRHRAGHHCQNAVIVI
                     LILAWEGIPRSLGDTLYQELTDTLRKYGNPTSRRCGLNDDRTCACQGKDPNTCGASFSF
                     GCSWSMYFNGCKYARSKTPRKFRLTGDNPKEEEVLRNSFQDLATEVAPLYKRLAPQAYQ
                     NQVTNEDVAIDCRLGLKEGRPFSGVTACMDFCAHAHKDQHNLYNGCTVVCTLTKEDNRC
                     VGQIPEDEQLHVLPLYKMASTDEFGSEENQNAKVSSGAIQVLTAFPREVRRLPEPAKSG
                     GGGSGGGGSGGGGSELWSDSEHNFLDENIGGVAVAPAHCSILIECARRELHATTPLKKP
                     NRCHPTRISLVFYQHKNLNQPNHGLALWEAKMKQLAERARQRQEEAARLG"
     protein_bind    complement(2924..2948)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2990..3576
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             3577..4284
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     polyA_signal    4364..4419
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   4867..5179
                     /label=LTR
                     /rpt_family="PiggyBAC LTR"
                     /note="LTR"
     promoter        complement(5324..5342)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5363..5379)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5387..5403)
                     /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(5411..5441)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5456..5477)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5765..6353)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6527..7384)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7385..7489)
                     /label=AmpR promoter