LOCUS       Exported                9835 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    1432 pcDNA3 flag MAGI1c
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9835)
  AUTHORS   Vazquez F, Grossman SR, Takahashi Y, Rokas MV, Nakamura N, Sellers 
            WR
  TITLE     Phosphorylation of the PTEN tail acts as an inhibitory switch by 
            preventing its recruitment into a protein complex.
  JOURNAL   J Biol Chem. 2001 Dec 28. 276(52):48627-30.
  PUBMED    11707428
REFERENCE   2  (bases 1 to 9835)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol 
            Chem. 2001 Dec 28. 276(52):48627-30."
FEATURES             Location/Qualifiers
     source          1..9835
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        173..552
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        553..756
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     707..727
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     801..820
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        801..819
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             845..868
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             872..895
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(5326..5344)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5327..5344)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5364..5381)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5370..5594
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5640..6068
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5727..5746)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5937..5958
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6077..6097)
                     /label=pBABE 3
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6082..6411
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6262..6397
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6324..6343
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6478..7272
                     /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(6532..6551)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7142..7161
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7446..7567
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7483..7502)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7537..7556
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7616..7632)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7616..7632)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7629..7651)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7640..7656
                     /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(7664..7694)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7709..7730
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7847..7864)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8018..8606)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8098..8117)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8777..9637)
                     /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     9400..9419
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9638..9742)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(9817..9835,1))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"