LOCUS       Exported                9333 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  expresses Wild type BRD4.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA5-Flag-BRD4-WT
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9333)
  AUTHORS   Shu S, Lin CY, He HH, Witwicki RM, Tabassum DP, Roberts JM, 
            Janiszewska M, Huh SJ, Liang Y, Ryan J, Doherty E, Mohammed H, Guo 
            H, Stover DG, Ekram MB, Peluffo G, Brown J, DSantos C, Krop IE, 
            Dillon D, McKeown M, Ott C, Qi J, Ni M, Rao PK, Duarte M, Wu SY, 
            Chiang CM, Anders L, Young RA, Winer EP, Letai A, Barry WT, Carroll 
            JS, Long HW, Brown M, Liu XS, Meyer CA, Bradner JE, Polyak K
  TITLE     Response and resistance to BET bromodomain inhibitors in 
            triple-negative breast cancer.
  JOURNAL   Nature. 2016 Jan 21;529(7586):413-7. doi: 10.1038/nature16508. Epub 
            2016 Jan 6.
  PUBMED    26735014
REFERENCE   2  (bases 1 to 9333)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nature16508"; journalName: "Nature"; date: "2016-01-21- 
            21"; volume: "529"; issue: "7586"; pages: "413-7"
FEATURES             Location/Qualifiers
     source          1..9333
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(23..42)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        214..593
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        594..797
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     748..768
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    799..817
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     848..872
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             971..994
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3212..3229
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5264..5281)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5270..5494
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5627..5646)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    5778..5825
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             5833..6855
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    6985..7106
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7022..7041)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7076..7095
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7155..7171)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7155..7171)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7168..7190)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7179..7195
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(7203..7233)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7248..7269
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7386..7403)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7557..8145)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7637..7656)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8316..9176)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8939..8958
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9177..9281)
                     /gene="bla"
                     /label=AmpR promoter