LOCUS       Exported               10411 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses V5- and EGFP-tagged human RRBP1 (with 54 decapeptide 
            repeats) in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA4 HisMax-V5-GFP-RRBP1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10411)
  AUTHORS   Hung V, Lam SS, Udeshi ND, Svinkina T, Guzman G, Mootha VK, Carr SA,
            Ting AY
  TITLE     Proteomic mapping of cytosol-facing outer mitochondrial and ER 
            membranes in living human cells by proximity biotinylation.
  JOURNAL   Elife. 2017 Apr 25;6. pii: e24463. doi: 10.7554/eLife.24463.
  PUBMED    28441135
REFERENCE   2  (bases 1 to 10411)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2017-04-25"; volume: "6. pii: e24463. doi"; pages: " 
            10.7554/eLife.24463"
FEATURES             Location/Qualifiers
     source          1..10411
                     /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             961..1002
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1012..1728
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1057..1078)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1318..1337)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1665..1686
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6419..6436)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6425..6649
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6695..7123
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6782..6801)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6992..7013
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7132..7152)
                     /label=pBABE 3
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7137..7466
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7317..7452
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7379..7398
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7514..7561
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7580..7954
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    8084..8205
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8121..8140)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8175..8194
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8254..8270)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8254..8270)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8267..8289)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8278..8294
                     /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(8302..8332)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8347..8368
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8485..8502)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8656..9244)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8736..8755)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(9415..10275)
                     /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     10038..10057
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10276..10380)
                     /gene="bla"
                     /label=AmpR promoter