LOCUS       Exported                8855 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCI-neo.mCherry-NEDD4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8855)
  AUTHORS   Nabhan JF, Pan H, Lu Q
  TITLE     Arrestin domain-containing protein 3 recruits the NEDD4 E3 ligase to
            mediate ubiquitination of the beta2-adrenergic receptor.
  JOURNAL   EMBO Rep. 2010 Aug;11(8):605-11. Epub 2010 Jun 18.
  PUBMED    20559325
REFERENCE   2  (bases 1 to 8855)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "EMBO Rep.";
            date: "2010-08"; volume: "11(8)"; pages: "605-11. Epub 2010 Jun 18"
FEATURES             Location/Qualifiers
     source          1..8855
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(10..27)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    45..166
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(56..75)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     110..129
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      352..807
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(439..458)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     649..670
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        924..1281
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(947..967)
                     /label=pBABE 3
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1132..1267
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1194..1213
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1332..2126
                     /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(1386..1405)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1996..2015
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2190..2238
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2279..2298)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2398..2420
                     /label=pGEX 3
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2458..2476)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2544..2648
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2649..3509
                     /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     complement(2867..2886)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3680..4268
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4169..4188
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4479..4858
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4859..5070
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5021..5041
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5067..5091
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          5231..5363
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     5346..5365
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        5408..5426
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5443..6150
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(5588..5606)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5856..5875
                     /label=mCherry-F
                     /note="mCherry, forward primer"