LOCUS       Exported               15084 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression of human ATM with Flag and His tags.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3.1(+)Flag-His-ATM wt
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15084)
  AUTHORS   Canman CE, Lim DS, Cimprich KA, Taya Y, Tamai K, Sakaguchi K, 
            Appella E, Kastan MB, Siliciano JD
  TITLE     Activation of the ATM kinase by ionizing radiation and 
            phosphorylation of p53.
  JOURNAL   Science. 1998 Sep 11;281(5383):1677-9.
  PUBMED    9733515
REFERENCE   2  (bases 1 to 15084)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science."; 
            date: "1998-09-11"; volume: "281(5383)"; pages: "1677-9"
FEATURES             Location/Qualifiers
     source          1..15084
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             984..1007
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1011..1028
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(10637..10655)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(10638..10655)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(10675..10692)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10681..10905
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10951..11379
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11038..11057)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11248..11269
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11388..11408)
                     /label=pBABE 3
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11393..11722
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11573..11708
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11635..11654
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             11789..12583
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(11843..11862)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     12453..12472
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    12757..12878
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12794..12813)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12848..12867
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12927..12943)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12927..12943)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12940..12962)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12951..12967
                     /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(12975..13005)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13020..13041
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13158..13175)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13329..13917)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13409..13428)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(14088..14948)
                     /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     14711..14730
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14949..15053)
                     /gene="bla"
                     /label=AmpR promoter