LOCUS       Exported                6880 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Flag tag with 3 residue spacer fused to the N-terminus of Lamp2b, a 
            transmembrane exosomal protein. 3 residue spacer then HA tag fused 
            to the C-terminus of Lamp2b..
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA Flag Lamp2b-HA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6880)
  AUTHORS   Hung ME, Leonard JN
  TITLE     Stabilization of exosome-targeting peptides via engineered 
            glycosylation.
  JOURNAL   J Biol Chem. 2015 Mar 27;290(13):8166-72. doi: 
            10.1074/jbc.M114.621383. Epub 2015 Feb 5.
  PUBMED    25657008
REFERENCE   2  (bases 1 to 6880)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1074/jbc.M114.621383"; journalName: "J Biol Chem"; date: 
            "2015-03-27- 27"; volume: "290"; issue: "13"; pages: "8166-72"
FEATURES             Location/Qualifiers
     source          1..6880
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(160..745)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(240..259)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(916..1776)
                     /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     1539..1558
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1777..1881)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1956..1975)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2147..2526
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2527..2730
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2681..2701
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2775..2794
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2775..2793
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2816..2825
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2906..2929
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4094..4120
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4217..4234)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4223..4447
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4493..4921
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4580..4599)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4790..4811
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4930..4950)
                     /label=pBABE 3
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4935..5264
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5115..5250
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5177..5196
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5313..6338
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    6468..6589
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6505..6524)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6559..6578
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6638..6654)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6638..6654)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6651..6673)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6662..6678
                     /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(6686..6716)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6731..6752
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(join(6869..6880,1..6))
                     /label=L4440
                     /note="L4440 vector, forward primer"